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1

Development of a bispecific monoclonal antibody for use in molecular hybridisation.  

PubMed

A mouse hybrid hybridoma (tetradoma) was prepared by fusing hybridomas producing monoclonal antibody to acetyl-aminofluorene with hybridomas producing antibody against calf intestine alkaline phosphatase. The tetradoma line established secreted immunoglobulin manifesting parental and bispecific binding characteristics. Bispecific monoclonal antibody was purified and used for a one-step immunodetection assay of non-radioactive DNA and RNA probes. The immunoassay developed was able to detect 5 pg DNA within 2 h and gave low background noise. PMID:8133070

Auriol, J; Guesdon, J L; Mazié, J C; Nato, F

1994-02-28

2

[Bispecific antibodies: what future?].  

PubMed

Monoclonal antibodies have emerged as a very successful class of therapeutic agents. In their native format, monoclonal antibodies are monospecific in that they recognize only one epitope, but their Fc domain also binds to FcfR-expressing cells. Attempts to improve the cytotoxicity of antibodies, particularly in the cancer field, have led to the design of bispecific antibodies: this can occur through various strategies, such as quadroma, thioether-linked Fab' gamma fragments or genetic engineering. Such bispecific antibodies have been developped to enhance immunotherapy, by bridging tumor cells and T cells, or radioimmunotherapy by combining bispecific antibodies and radiolabeled bivalent haptens that bind cooperatively to target cells. Multiple further applications can be envisaged such as targeting two different antigens on the same cell, or two epitopes of the same antigen. Although progresses have been slowed by technical constraints, there is little doubt that this class of novel antibodies derivatives will experience a promising development. PMID:20035697

Pèlegrin, André; Robert, Bruno

2009-12-01

3

Antitumor immunity: easy as 1, 2, 3 with monoclonal bispecific trifunctional antibodies?  

PubMed

Monoclonal antibodies occupy an increasing niche in the arsenal available to treat cancer. Several developments have rendered this the fastest growing sector in the pharmaceutical industry. Traditionally, antibodies were developed to block key signaling molecules implicated in tumor progression. However, antibodies also recruit additional immune effector mechanisms against tumors, a property that may be exploited for clinical benefit. Bispecific antibodies represent one such strategy in which elements derived from two monoclonal antibodies are incorporated into a single molecular species. Commonly, the bispecific approach is used to achieve simultaneous cross-linking of CD3 and a tumor antigen such as epithelial cell adhesion molecule (EpCAM), thereby recruiting T-cell activation to the tumor cell surface. A further sophistication involves the engineering of trifunctional derivatives such as the clinically approved agent, catumaxomab. Catumaxomab has antigen-binding arms that engage CD3 and EpCAM and a constant domain that recruits Fc receptor-bearing cells, notably monocytes, dendritic cells, and natural killer cells. Owing to this triangular binding capability, catumaxomab can activate both innate and adaptive immune effector mechanisms in addition to promoting immunologic memory. Recent data indicate that this agent can also promote immunogenic cell death, particularly when used in combination with selected chemotherapeutic agents such as oxaliplatin. PMID:24014596

Maher, John; Adami, Antonella A

2013-09-15

4

Development of a bispecific monoclonal antibody against a gallium-67 chelate and the human melanoma-associated antigen p97 for potential use in pretargeted immunoscintigraphy  

Microsoft Academic Search

A bispecific monoclonal antibody (bsmAb) has been developed against the human melanoma-associated antigen p97 and an octahedral gallium chelate (Ga-HBED) using the hybrid hybridoma technology. As tetradomas were expected to produce a maximum of ten different molecular species of immunoglobulins, the bispecific antibody was purified from this mixture by consecutive protein A affinity and cation-exchange chromatographic techniques. Although it was

Chezian Somasundaram; Siegfried Matzku; Jochen Schuhmacher; Margot Zöller

1993-01-01

5

Bispecific Antibodies for Diagnostic Applications  

Microsoft Academic Search

\\u000a Bispecific monoclonal antibodies (BsMAb) are unique engineered macromolecules that have two different pre-determined binding\\u000a specificities. Their ability to simultaneously bind to a specific antigen and a given detection moiety enables them to function\\u000a as excellent bifunctional immunoprobes in diagnostic assays. BsMAb are being exploited for the development of simple, rapid,\\u000a and highly sensitive immunoassays for diagnosis of bacterial and viral

Archana Parashar; Susmita Sarkar; Advaita Ganguly; Sai Kiran Sharma; Mavanur R. Suresh

6

Bispecific Antibodies: Molecules That Enable Novel Therapeutic Strategies  

Microsoft Academic Search

Bispecific antibodies are unique in the sense that they can bind simultaneously two different antigens. This property enables the development of therapeutic strategies that are not possible with conventional monoclonal antibodies. The large panel of imaginative bispecific antibody formats that has been developed reflects the strong interest for these molecules. Although in many cases the manufacturing of clinical grade material

Nicolas Fischer; Olivier Léger

2007-01-01

7

Dual targeting strategies with bispecific antibodies  

PubMed Central

Monoclonal antibodies are widely used for the treatment of cancer, inflammatory and infectious diseases and other disorders. Most of the marketed antibodies are monospecific and therefore capable of interacting and interfering with a single target. However, complex diseases are often multifactorial in nature, and involve redundant or synergistic action of disease mediators or upregulation of different receptors, including crosstalk between their signaling networks. Consequently, blockade of multiple, different pathological factors and pathways may result in improved therapeutic efficacy. This result can be achieved by combining different drugs, or use of the dual targeting strategies applying bispecific antibodies that have emerged as an alternative to combination therapy. This review discusses the various dual targeting strategies for which bispecific antibodies have been developed and provides an overview of the established bispecific antibody formats.

2012-01-01

8

A revival of bispecific antibodies  

Microsoft Academic Search

Bispecific antibodies usually do not occur in nature but are constructed by recombinant DNA or cell-fusion technologies. Most are designed to recruit cytotoxic effector cells of the immune system effectively against pathogenic target cells. This complex task explains why, after more than 15 years of extensive research, many different formats of bispecific antibodies have been developed but only a few

Peter Kufer; Ralf Lutterbüse; Patrick A. Baeuerle

2004-01-01

9

Local but no systemic immunomodulation by intraperitoneal treatment of advanced ovarian cancer with autologous T lymphocytes re-targeted by a bi-specific monoclonal antibody  

Microsoft Academic Search

We have reported a 27% overall anti-tumor response using i.p. immunotherapy of advanced ovarian carcinoma with autologous, ex vivo expanded, T lymphocytes re-targeted with bi-specific monoclonal antibody OC\\/TR, combined with soluble OC\\/TR and low-dose recombinant interleukin-2 (IL-2). This treatment had no effect on extraperitoneal disease. Therefore we studied in 13 patients whether this immunotherapeutic protocol resulted only in local or

Cor H. J. Lamers; Reinder L. H. Bolhuis; Sven O. Warnaar; Gerrit Stoter; Jan W. Gratama

1997-01-01

10

Bispecific antibodies for cancer therapy  

PubMed Central

With 23 approvals in the US and other countries and four approvals outside US, antibodies are now widely recognized as therapeutic molecules. The therapeutic and commercial successes met by rituximab, trastuzumab, cetuximab and other mAbs have inspired antibody engineers to improve the efficacy of these molecules. Consequently, a new wave of antibodies with engineered Fc leading to much higher effector functions such as antibody-dependent cell-mediated cytotoxicity or complement-dependent cytotoxicity is being evaluated in the clinic, and several approvals are expected soon. In addition, research on a different class of antibody therapeutics, bispecific antibodies, has recently led to outstanding clinical results, and the first approval of the bispecific antibody catumaxomab, a T cell retargeting agent that was approved in the European Union in April 2009. This review describes the most recent advances and clinical study results in the field of bispecific antibodies, a new class of molecules that might outshine conventional mAbs as cancer immunotherapeutics in a near future.

Baty, Daniel

2009-01-01

11

Bispecific Antibodies from Hybrid Hybridoma  

Microsoft Academic Search

\\u000a Hybrid hybridomas (also termed quadromas or tetradomas) are man-made cell lines that secrete bispecific antibodies (bsAb)\\u000a with two different specificities being able to crosslink two distinct molecules. Such antibodies do not occur in nature and\\u000a have been originally developed to improve immunohistochemical staining procedures and immunoassays (Milstein and Cuello 1983;\\u000a Suresh et al. 1986). Interestingly, the fusion of two immunoglobulin-producing

Gerhard Moldenhauer

12

Cancer therapy with bispecific antibodies: Clinical experience  

PubMed Central

The binding of at least two molecular targets simultaneously with a single bispecific antibody is an attractive concept. The use of bispecific antibodies as possible therapeutic agents for cancer treatment was proposed in the mid-1980s. The design and production of bispecific antibodies using antibody- and/or receptor-based platform technology has improved significantly with advances in the knowledge of molecular manipulations, protein engineering techniques, and the expression of antigens and receptors on healthy and malignant cells. The common strategy for making bispecific antibodies involves combining the variable domains of the desired mAbs into a single bispecific structure. Many different formats of bispecific antibodies have been generated within the research field of bispecific immunotherapeutics, including the chemical heteroconjugation of two complete molecules or fragments of mAbs, quadromas, F(ab’)2, diabodies, tandem diabodies and single-chain antibodies. This review describes key modifications in the development of bispecific antibodies that can improve their efficacy and stability, and provides a clinical perspective on the application of bispecific antibodies for the treatment of solid and liquid tumors, including the promises and research limitations of this approach.

Thakur, Archana; Lum, Lawrence G

2013-01-01

13

Monoclonal Antibodies.  

ERIC Educational Resources Information Center

Monoclonal antibodies have provided an exciting addition to the "armory" of the molecular biologist and immunologist. This article discusses briefly the concept of, techniques available for, production of, and possible uses of monoclonal antibodies. (Author)

Killington, R. A.; Powell, K. L.

1984-01-01

14

Two distinct monoclonal antibodies raised against mouse beta nerve growth factor. Generation of bi-specific anti-nerve growth factor anti-horseradish peroxidase antibodies for use in a homogeneous enzyme immunoassay.  

PubMed

Two hybridomas producing monoclonal antibodies against mouse beta nerve growth factor (NGF) were obtained from the fusion of hyperimmune splenocytes from rats immunized with polymerized beta-NGF and Sp2/0.Ag mouse myeloma cells. The monoclonal antibodies coded IgG 24 and 30 produced and secreted by the hybrid cells are both of the IgG2a subclass. Both monoclonal antibodies are capable of recognizing native NGF coated on microassay plates as well as the denatured factor on Western blots. However, only IgG 30 has been found to block NGF-induced process outgrowth from the rat pheochromocytoma cell line (PC12) as well as NGF-induced increase in choline acetyltransferase activity in rat primary septal cell cultures. In addition, only IgG 30 was able to detect immunocytochemically NGF-immunoreactive sites in fixed tissue. And, finally, IgG 24 could not compete for IgG 30 binding to immobilized native NGF. Consequently, it appears that these antibodies are recognizing different epitopes on the NGF molecule. Neither monoclonal antibody displayed any crossreactivity with serum albumin, aprotinin, epidermal growth factor or insulin. A hybrid-hybridoma producing bi-specific anti-NGF anti-horseradish peroxidase (HRP) monoclonal antibodies was generated from the fusion of an azaguanine resistant anti-HRP hybridoma, coded RAP2.Ag and the anti-NGF IgG 30 hybridoma treated with emetine. The potential merits of using these bi-specific antibodies in combination with their mono-specific anti-NGF parent in a homogeneous sandwich immunoassay for the quantitation of NGF are discussed. PMID:1999653

Kenigsberg, R L; Elliott, P J; Cuello, A C

1991-02-15

15

Time resolved native ion-mobility mass spectrometry to monitor dynamics of IgG4 Fab arm exchange and "bispecific" monoclonal antibody formation.  

PubMed

Monoclonal antibodies (mAbs) and derivatives such as antibody-drug conjugates (ADC) and bispecific antibodies (bsAb), are the fastest growing class of human therapeutics. Most of the therapeutic antibodies currently on the market and in clinical trials are chimeric, humanized, and human immunoglobulin G1 (IgG1). An increasing number of IgG2s and IgG4s that have distinct structural and functional properties are also investigated to develop products that lack or have diminished antibody effector functions compared to IgG1. Importantly, wild type IgG4 has been shown to form half molecules (one heavy chain and one light chain) that lack interheavy chain disulfide bonds and form intrachain disulfide bonds. Moreover, IgG4 undergoes a process of Fab-arm exchange (FAE) in which the heavy chains of antibodies of different specificities can dissociate and recombine in bispecific antibodies both in vitro and in vivo. Here, native mass spectrometry (MS) and time-resolved traveling wave ion mobility MS (TWIM-MS) were used for the first time for online monitoring of FAE and bsAb formation using Hz6F4-2v3 and natalizumab, two humanized IgG4s which bind to human Junctional Adhesion Molecule-A (JAM-A) and alpha4 integrin, respectively. In addition, native MS analysis of bsAb/JAM-A immune complexes revealed that bsAb can bind up to two antigen molecules, confirming that the Hz6F4 family preferentially binds dimeric JAM-A. Our results illustrate how IM-MS can rapidly assess bsAb structural heterogeneity and be easily implemented into MS workflows for bsAb production follow up and bsAb/antigen complex characterization. Altogether, these results provide new MS-based methodologies for in-depth FAE and bsAb formation monitoring. Native MS and IM-MS will play an increasing role in next generation biopharmaceutical product characterization like bsAbs, antibody mixtures, and antibody-drug conjugates (ADC) as well as for biosimilar and biobetter antibodies. PMID:24007193

Debaene, François; Wagner-Rousset, Elsa; Colas, Olivier; Ayoub, Daniel; Corvaïa, Nathalie; Van Dorsselaer, Alain; Beck, Alain; Cianférani, Sarah

2013-10-15

16

In vitro and in vivo stability and anti-tumour efficacy of an anti-EGFR/anti-CD3 F(ab')2 bispecific monoclonal antibody.  

PubMed Central

The in vitro and in vivo stability and anti-tumour efficacy of the anti-EGFR/anti-CD3 bispecific monoclonal antibody (biMAb), M26.1, were analysed. The interaction of the intact biMAb with Fc receptor I (Fc gamma RI) present on human leucocytes was not observed when the antibody was used as an F(ab')2 fragment. A CD8+ T-cell clone coated with M26.1 F(ab')2 was as effective as the intact biMAb in inducing IGROV1 target cell lysis when tested in a 51Cr-release assay. Variable levels of reduction of F(ab')2 to monovalent F(ab') were observed upon incubation with human ovarian cancer ascitic fluid (OCAF) or with human glioblastoma cavity fluid (GCF), but not with mouse or human sera. Activated lymphocytes coated with F(ab')2 and incubated in vitro with GCF or OCAF for 24 and 48 h respectively maintained their targeting. Thus, the F(ab')2, when present as a soluble molecule, but not when bound to T cells, might lose some functional activity as a consequence of partial reduction to F(ab'). In normal mice, M26.1 F(ab')2 retained full cytotoxic activity in the circulation, and clearance values were similar to those obtained with parental and other MAb F(ab')2. Treatment of IGROV1 tumour-bearing mice with activated human lymphocytes coated with the M26.1 F(ab')2 significantly prolonged survival of the animals compared with tumour-bearing untreated and control mice treated with lymphocytes or F(ab')2 alone. Together, these results suggest the clinical usefulness of bispecific M26.1 F(ab')2 as a targeting agent for local treatment of tumours such as glioma and ovarian cancers that express variable levels of epidermal growth factor receptor (EGFR).

Negri, D. R.; Tosi, E.; Valota, O.; Ferrini, S.; Cambiaggi, A.; Sforzini, S.; Silvani, A.; Ruffini, P. A.; Colnaghi, M. I.; Canevari, S.

1995-01-01

17

Targeting of saporin to CD25-positive normal and neoplastic lymphocytes by an anti-saporin/anti-CD25 bispecific monoclonal antibody: in vitro evaluation.  

PubMed Central

This study has been designed to verify the specific toxicity of saporin, a type 1 ribosome-inactivating protein (RIP), with the same activity as ricin A chain, targeted by a bispecific monoclonal antibody (bimAb) recognising both the CD25 antigen and the RIP. The CD25 antigen is expressed by lymphoid populations upon activation and by leukaemias and lymphomas with an activated membrane phenotype (Hodgkin's lymphoma, anaplastic large cell lymphoma, adult T cell leukaemia). The bimAb-saporin mixture was tested on CD25+ targets at different bimAb and saporin concentrations. Saporin, in the presence of a bimAb concentration of 10(-9) M, inhibited protein synthesis by CD25+ neoplastic lymphocytes (L540 and MT2 cell lines) with IC50S (concentrations giving 50% of inhibition) ranging from 8 x 10(-12) M to 3 x 10(-11) M. The saporin-bimAb mixture was also effective in blocking the phytohaemagglutinin-driven proliferation of normal lymphocytes, whereas it displayed the same level of toxicity exerted by saporin alone on an irrelevant CD25-negative cell line (EBV-infected B lymphoblastoid cell line). From these results it is possible to envisage a clinical use of this bimAb as a cytotoxic agent for CD25+ leukaemias and lymphomas, as well as an immunosuppressive agent for severe immune disorders such as graft-vs-host disease (GVHD) and transplanted organ rejection.

Tazzari, P. L.; Zhang, S.; Chen, Q.; Sforzini, S.; Bolognesi, A.; Stirpe, F.; Xie, H.; Moretta, A.; Ferrini, S.

1993-01-01

18

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies  

PubMed Central

We demonstrate here a rapid alternative method for the production of functional bi-specific antibodies using the mild reducing agent 2-mercaptoethanesulfonic acid sodium salt (MESNA). Following reduction of a mixture of two monoclonal antibodies with MESNA to break inter heavy chain bonds, this solution is dialysed under oxidising conditions and antibodies are allowed to reform. During this reaction a mixture of antibodies is formed, including parental antibodies and bi-specific antibody. Bi-specific antibodies are purified over two sequential affinity columns. Following purification, bi-specificity of antibodies is determined in enzyme-linked immunosorbent assays and by flow cytometry. Using this redox method we have been successful in producing hybrid and same-species bi-specific antibodies in a time frame of 6–10 working days, making this production method a time saving alternative to the time-consuming traditional heterohybridoma technology for the production of bi-specific antibodies for use in early pilot studies. The use of both rat and mouse IgG antibodies forming a rat/mouse bi-specific antibody as well as producing a pure mouse bi-specific antibody and a pure rat bi-specific antibody demonstrates the flexibility of this production method.

Carlring, Jennifer; De Leenheer, Evy; Heath, Andrew William

2011-01-01

19

World Bispecific Antibody Summit, September 27-28, 2011, Boston, MA  

PubMed Central

With more than 30 therapeutic monoclonal antibodies (mAbs) approved and annual global sales of the products at ?$50 billion in 2010, these products have proven to be successful in many ways. Nevertheless, there is room for improvement in performance, and substantial unmet medical needs remain. As a consequence, numerous organizations are devoting resources to engineering novel mAbs such as bispecific antibodies that have increased functionality compared with unmodified IgG molecules. The World Bispecific Antibody Summit, organized by Hanson Wade, drew over 100 participants to Boston to discuss engineering novel bispecific antibodies, generating lead candidates and clinical study and commercialization of the molecules. Approaches such as the trifunctional antibody (TRION), dual variable domain-Ig (Abbott), two-in-one (Genentech), dual affinity retargeting (MacroGenics), kappa-lambda body (NovImmune), bispecific T-cell engager (Micromet) and chemical generation (CovX/Pfizer) were discussed in detail. In addition, posters describing bispecific Affibody® molecules for targeting of EGFR and HER2 (Affibody), T-cell receptor based bi-specifics that target HLA-peptides (Immunocore), a novel mAb-Fv bispecific antibody format utilizing Fc region (Xencore), generation of a tetravalent bispecific antibody against IL4 and IL13 for the treatment of idiopathic pulmonary fibrosis (Sanofi), Combining Affibody® molecules and the AlbumodTM technology to create long acting multispecific protein therapeutics (Royal Institute of Technology, Affibody) and COVA301 as a highly potent bispecific inhibitor of IL-17A and TNF? (Covagen) were presented.

Dhimolea, Eugen

2012-01-01

20

World Bispecific Antibody Summit, September 27-28, 2011, Boston, MA.  

PubMed

With more than 30 therapeutic monoclonal antibodies (mAbs) approved and annual global sales of the products at ~$50 billion in 2010, these products have proven to be successful in many ways. Nevertheless, there is room for improvement in performance, and substantial unmet medical needs remain. As a consequence, numerous organizations are devoting resources to engineering novel mAbs such as bispecific antibodies that have increased functionality compared with unmodified IgG molecules. The World Bispecific Antibody Summit, organized by Hanson Wade, drew over 100 participants to Boston to discuss engineering novel bispecific antibodies, generating lead candidates and clinical study and commercialization of the molecules. Approaches such as the trifunctional antibody (TRION), dual variable domain-Ig (Abbott), two-in-one (Genentech), dual affinity retargeting (MacroGenics), kappa-lambda body (NovImmune), bispecific T-cell engager (Micromet) and chemical generation (CovX/Pfizer) were discussed in detail. In addition, posters describing bispecific Affibody® molecules for targeting of EGFR and HER2 (Affibody), T-cell receptor based bi-specifics that target HLA-peptides (Immunocore), a novel mAb-Fv bispecific antibody format utilizing Fc region (Xencore), generation of a tetravalent bispecific antibody against IL4 and IL13 for the treatment of idiopathic pulmonary fibrosis (Sanofi), Combining Affibody® molecules and the Albumod™ technology to create long acting multispecific protein therapeutics (Royal Institute of Technology, Affibody) and COVA301 as a highly potent bispecific inhibitor of IL-17A and TNF-? (Covagen) were presented. PMID:22327426

Dhimolea, Eugen; Reichert, Janice M

2012-01-01

21

Universal Bispecific Antibody for Targeting Tumor Cells for Destruction by Cytotoxic T Cells  

NASA Astrophysics Data System (ADS)

Previous studies have demonstrated that bispecific hybrid antibodies produced by cell-cell fusion or chemically conjugated heteroaggregates can direct cytotoxic T lymphocytes to kill target cells for which they have no intrinsic specificity, a phenomenon we call effector cell retargeting (ECR). These studies used bispecific reagents with one specificity directed to CD3 or Ti on the effector cell and the other directed to a target cell antigen. To avoid the need to create different hybrid hybridomas for each target antigen we have developed a universal means to elicit ECR through the use of an antiglobulin step. We have constructed a bispecific hybrid antibody with dual specificity for CD3 and a rat immunoglobulin light chain allotype. This bispecific antibody could mediate ECR to a range of target cells, each coated with distinct surface-binding rat monoclonal antibodies. A particular advantage of targeting to surface-bound monoclonal antibodies is that all other available effector systems may also attack the same antibody-coated target cell.

Gilliland, Lisa K.; Clark, Michael R.; Waldmann, Herman

1988-10-01

22

Generation of bispecific IgG antibodies by structure-based design of an orthogonal Fab interface.  

PubMed

Robust generation of IgG bispecific antibodies has been a long-standing challenge. Existing methods require extensive engineering of each individual antibody, discovery of common light chains, or complex and laborious biochemical processing. Here we combine computational and rational design approaches with experimental structural validation to generate antibody heavy and light chains with orthogonal Fab interfaces. Parental monoclonal antibodies incorporating these interfaces, when simultaneously co-expressed, assemble into bispecific IgG with improved heavy chain-light chain pairing. Bispecific IgGs generated with this approach exhibit pharmacokinetic and other desirable properties of native IgG, but bind target antigens monovalently. As such, these bispecific reagents may be useful in many biotechnological applications. PMID:24463572

Lewis, Steven M; Wu, Xiufeng; Pustilnik, Anna; Sereno, Arlene; Huang, Flora; Rick, Heather L; Guntas, Gurkan; Leaver-Fay, Andrew; Smith, Eric M; Ho, Carolyn; Hansen-Estruch, Christophe; Chamberlain, Aaron K; Truhlar, Stephanie M; Conner, Elaine M; Atwell, Shane; Kuhlman, Brian; Demarest, Stephen J

2014-02-01

23

Monoclonal antibodies and immobilized antibodies  

Microsoft Academic Search

Antibodies in both their free and immobilized state have been the object of considerable industrial and academic interest.\\u000a A variety of methods are used for preparing and immobilizing antibodies. Applications for monoclonal antibodies include the\\u000a preparation of therapeutics, diagnostics, and in affinity fractionation. Recent US patents on monoclonal and immobilized antibodies\\u000a and scientific literature on monoclonal antibodies are surveyed. A

Robert J. Linhardt; C. W. Abell; R. M. Denney; B. W. Altrock; R. Auerbach; S. D. Bernal; R. E. Canfield; P. H. Ehrlich; W. R. Moyle; T. S. Chan; T. W. Chang; N. T. Chang; J. A. Cidlowski; M. D. Viceps; R. J. Cote; D. M. Morrissey; A. N. Houghton; E. J. Beattie; H. F. Oettgen; L. J. Old; C. M. Croce; R. S. Cubicciotti; A. E. Karu; R. M. Krauss; J. S. Cullor; A. Deutsch; H. Brandwein; H. Platt; D. M. Hunter; A. Dubitsky; S. M. Durham; F. A. Dolbeare; J. W. Gray; G. R. Dreesman; C. E. Kendall; J. C. Egrie; A. R. Frackelton; H. N. Eisen; A. H. Ross; S. Gay; G. Geirnaert; J. E. Geltosky; E. H. Goldberg; E. Goldwasser; C. Kavinsky; T. L. Weiss; H. G. Gratzner; B. Hampar; M. Zweig; S. D. Showalter; H. H. Handley; M. C. Glassy; Y. Hagiwara; H. Hagiwara; C. M. Huang; S. N. Cohen; J. V. Hughes; E. M. Scolnick; J. E. Tomassini; R. Jefferis; J. Steensgaard; H. S. Kaplan; N. N. H. Teng; K. S. Earn; R. F. Calvo; L. Kass; J. R. Kettman; M. V. Norgard; M. B. Khazaeli; W. H. Beierwaltes; B. G. England; P. C. Kung; G. Goldstein; L. Lanier; J. Phillips; N. L. Warner; J. W. Larrick; A. R. Raubitschek; K. E. Truitt; H. Lazarus; J. F. Schwaber; J. Lewicki; C. Lewis; J. V. Olander; W. R. Tolbert; E. L. Milford; C. B. Carpenter; J. M. Paradysz; D. F. Mosher; J. L. Mulshine; J. D. Minna; K. A. Murray; D. M. Neville; R. J. Youle; M. Nicolson; I. Pastan; M. C. Willingham; D. J. Fitzgerald; A. Pucci; A. M. Smithyman; M. B. Slade; P. W. French; G. Wijffels; C. S. Pukel; K. O. Lloyd; L. R. Travassos; W. G. Dippold; R. P. Reckel; J. L. Harris; R. Wellerson; S. M. Shaw; P. M. Kaplan; E. L. Reinherz; S. F. Schlossman; S. C. Mener; J. Sakamoto; C. C. Cordon; E. Friedman; C. L. Finstad; W. E. Enker; M. R. Melamed; J. F. Oettgen; P. J. Scannon; L. E. Spitler; H. M. Lee; R. T. Kawahata; R. P. Mischak; J. Schlom; D. Colcher; M. Nuti; P. H. Hand; F. Austin; G. D. Shockman; D. E. Jackson; W. Wong; Z. Steplewski; H. Koprowski; M. Herlyn; M. Strand; I. S. Trowbridge; D. L. Urdal; C. J. March; S. K. Dower; J. R. Wands; V. R. Zurawski; C. A. White; R. Dulbecco; W. R. Allen; E. C. Arnold; M. Flasher; H. H. Freedman; T. D. Heath; P. Shek; D. Papahadjopoulos; M. Ikeda; S. Sakamoto; K. Suzuki; M. Kuboyama; Y. Harada; A. Kawashiri; E. Takahashi; H. S. Lee; S. Margel; R. C. Nowinski; A. S. Hoffman; J. W. Peterson; K. B. Platt; D. E. Reed; F. X. Real; M. J. Mattes; P. O. Livingston; A. Rembaum; R. C. K. Yen; R. Rosenstein; B. Schneider

1987-01-01

24

Synthesis of bispecific antibodies using genetically encoded unnatural amino acids.  

PubMed

Bispecific antibodies were constructed using genetically encoded unnatural amino acids with orthogonal chemical reactivity. A two-step process afforded homogeneous products in excellent yield. Using this approach, we synthesized an anti-HER2/anti-CD3 bispecific antibody, which efficiently cross-linked HER2+ cells and CD3+ cells. In vitro effector-cell mediated cytotoxicity was observed at picomolar concentrations. PMID:22642368

Kim, Chan Hyuk; Axup, Jun Y; Dubrovska, Anna; Kazane, Stephanie A; Hutchins, Benjamin A; Wold, Erik D; Smider, Vaughn V; Schultz, Peter G

2012-06-20

25

A cell-penetrating bispecific antibody for therapeutic regulation of intracellular targets.  

PubMed

The therapeutic use of antibodies is restricted by the limited access of antibodies to intracellular compartments. To overcome this limitation, we developed a cell-penetrating monoclonal antibody, mAb 3E10, as an intracellular delivery vehicle for the intracellular and intranuclear delivery of antibodies constructed as bispecific single-chain Fv fragments. Because MDM2 is an important target in cancer therapy, we selected monoclonal antibody (mAb) 3G5 for intracellular transport. mAb 3G5 binds MDM2 and blocks binding of MDM2 to p53. Here, we show that the resulting 3E10-3G5 bispecific antibody retains cell-penetrating and MDM2-binding activity, increases tumor p53 levels, and inhibits growth of MDM2-addicted tumors. The use of cell-penetrating bispecific antibodies in targeted molecular therapy will significantly broaden the spectrum of accessible intracellular targets and may have a profound impact in cancer therapy. PMID:22863609

Weisbart, Richard H; Gera, Joseph F; Chan, Grace; Hansen, James E; Li, Erica; Cloninger, Cheri; Levine, Arnold J; Nishimura, Robert N

2012-10-01

26

Targeting T Cells with Bispecific Antibodies for Cancer Therapy  

PubMed Central

Bispecific antibodies (BiAbs) offer a unique opportunity to redirect immune effector cells to kill cancer cells. BiAbs combine the benefits of different binding specificities of two monoclonal antibodies (mAbs) into a single construct. This unique feature of BiAbs enables approaches that are not possible with single mAbs. Advances in antibody engineering and antigen profiling of malignant cells have led to the development of a number of BiAb formats and their combinations for redirecting effector cells to tumor targets. There have been significant advances in the design and application of BiAbs for intravenous and local injection. The initial barrier of cytokine storm has been partially overcome by more recent constructs that have improved clinical effectiveness without dose-limiting toxicities. Since the recent revival of BiAbs, there has been multiple, ongoing, phase I/II and III trials, and some promising clinical outcomes have been reported in completed clinical studies. This review focuses on arming T cells with BiAbs to create the ‘poor man's cytotoxic lymphocyte’.

Lum, Lawrence G.; Thakur, Archana

2013-01-01

27

From whole monoclonal antibodies to single domain antibodies: think small.  

PubMed

The development of therapeutic monoclonal antibodies over the last 35 years has led to the emergence of a new class of useful therapeutic molecules. These "first generation" antibodies have been obtained thanks to the conjugated and huge efforts of both academic and biotech researchers. About 30 monoclonal antibodies are currently approved for therapeutic use in Europe, USA, and China. Strikingly, only a restricted number of these antibodies are immunoglobulin fragments, single variable domains, or multiunit formats based on the engineering of immunoglobulin variable domains. In the present chapter, we will review the major steps of the therapeutic antibodies history and we will highlight the enormous potential of antibody fragments, either used as multiunits such as bispecific antibodies, single units, or as cell modifiers such as intrabodies or cell surface-expressed molecules. PMID:22886242

Teillaud, Jean-Luc

2012-01-01

28

Quantification of cell surface proteins with bispecific antibodies  

PubMed Central

Flow cytometry is an established method for fast and accurate quantitation of cellular protein levels and requires fluorescently labeled antibodies as well as calibration standards. A critical step for quantitation remains the production of suitable detection antibodies with a precisely defined ratio of antigen-binding sites to fluorophores. Problems often arise as a consequence of inefficient and unspecific labeling which can influence antibody properties. In addition, the number of incorporated fluorophores necessitates a special normalization step for quantitation. To address these problems, we constructed different mono- and bivalent bispecific antibodies with binding site(s) for the cell surface antigens, cMET, EGFR1/HER1, ErbB2/HER2 or ErbB3/HER3 and with an additional digoxigenin-binding single-chain Fv fusion. The fluorophore Cy5 was covalently coupled to digoxigenin and quantitatively bound by the bispecific antibody. A panel of tumor cell lines was assessed under different culture conditions for absolute receptor expression levels of the indicated antigens and the data were set in relation to mRNA, gene count and immunoblot data. We could reproducibly quantify these receptors, omit the otherwise required normalization step and demonstrate the superiority of a 1 + 1 bispecific antibody. The same antibodies were also used to quantify the number of proteins in intracellular vesicles in confocal microscopy. The antibodies can be stored like regular antibodies and can be coupled with different digoxigenin-labeled fluorophores which makes them excellent tools for FACS and imaging-based experiments.

Panke, C.; Weininger, D.; Haas, A.; Schelter, F.; Schlothauer, T.; Bader, S.; Sircar, R.; Josel, H.P.; Baer, U.; Burtscher, H.; Mundigl, O.; Grote, M.; Brinkmann, U.; Sustmann, C.

2013-01-01

29

Bispecific T-cell engaging antibodies for cancer therapy.  

PubMed

There is increasing evidence that T cells are able to control tumor growth and survival in cancer patients, both in early and late stages of the disease. However, tumor-specific T-cell responses are difficult to mount and sustain in cancer patients, and are limited by numerous immune escape mechanisms of tumor cells selected during immunoediting. An alternative approach to engage T cells for cancer therapy are antibodies, which are bispecific for a surface target antigen on cancer cells, and for CD3 on T cells. These are capable of connecting any kind of cytotoxic T cell to a cancer cell, independently of T-cell receptor specificity, costimulation, or peptide antigen presentation. Here, we review the principle of a new class of bispecific antibodies called BiTE (for "bispecific T-cell engager") antibodies. Recent results from clinical studies with a CD19/CD3-bispecific BiTE antibody suggest that this therapeutic paradigm is finally showing promise for treatment of both bulky and minimal residual disease. PMID:19509221

Baeuerle, Patrick A; Reinhardt, Carsten

2009-06-15

30

Monoclonal antibody "gold rush".  

PubMed

The market, sales and regulatory approval of new human medicines, during the past few years, indicates increasing number and share of new biologics and emergence of new multibillion dollar molecules. The global sale of monoclonal antibodies in 2006 were $20.6 billion. Remicade had annual sales gain of $1 billion during the past 3 years and five brands had similar increase in 2006. Rituxan with 2006 sales of $4.7 billion was the best selling monoclonal antibody and biological product and the 6th among the top selling medicinal brand. It may be the first biologic and monoclonal antibody to reach $10 billion annual sales in the near future. The strong demand from cancer and arthritis patients has surpassed almost all commercial market research reports and sales forecast. Seven monoclonal antibody brands in 2006 had sales exceeding $1 billion. Humanized or fully human monoclonal antibodies with low immunogenicity, enhanced antigen binding and reduced cellular toxicity provide better clinical efficacy. The higher technical and clinical success rate, overcoming of technical hurdles in large scale manufacturing, low cost of market entry and IND filing, use of fully human and humanized monoclonal antibodies has attracted funds and resources towards R&D. Review of industry research pipeline and sales data during the past 3 years indicate a real paradigm shift in industrial R&D from pharmaceutical to biologics and monoclonal antibodies. The antibody bandwagon has been joined by 200 companies with hundreds of new projects and targets and has attracted billions of dollars in R&D investment, acquisitions and licensing deals leading to the current Monoclonal Antibody Gold Rush. PMID:17691940

Maggon, Krishan

2007-01-01

31

Monoclonal antibodies and cancer therapy  

SciTech Connect

These proceedings collect papers on the subject of monoclonal antibodies. Topics include: Monoclonal antibody, biochemical effects and cancer therapeutic potential of tunicamycin, use of monoclonal antibodies for detection of lymph node metastases, active specific immunotherapy, and applications of monoclonal antibodies to investigations of growth factors.

Reisfeld, R.A.; Sell, S.

1985-01-01

32

Targeting T cells to tumor cells using bispecific antibodies.  

PubMed

The immune system, and in particular T cells, can be harnessed to treat cancer. Several bispecific T cell engaging antibodies of the BiTE® format are in early or late-stage clinical development. These small recombinant antibody constructs effectively trigger killing of cancer cells by temporarily attached, polyclonal T cells. Blinatumomab, a CD19/CD3-bispecific BiTE® antibody, has demonstrated high clinical activity in B cell leukemia and lymphoma patients. Three additional BiTE antibodies directed against surface target antigen expressed on solid tumors are being evaluated in phase I clinical trials. Alternative approaches to direct polyclonal T cells to kill cancer cells are under intense investigation. PMID:23623807

Frankel, Stanley R; Baeuerle, Patrick A

2013-06-01

33

Quantification of cell surface proteins with bispecific antibodies.  

PubMed

Flow cytometry is an established method for fast and accurate quantitation of cellular protein levels and requires fluorescently labeled antibodies as well as calibration standards. A critical step for quantitation remains the production of suitable detection antibodies with a precisely defined ratio of antigen-binding sites to fluorophores. Problems often arise as a consequence of inefficient and unspecific labeling which can influence antibody properties. In addition, the number of incorporated fluorophores necessitates a special normalization step for quantitation. To address these problems, we constructed different mono- and bivalent bispecific antibodies with binding site(s) for the cell surface antigens, cMET, EGFR1/HER1, ErbB2/HER2 or ErbB3/HER3 and with an additional digoxigenin-binding single-chain Fv fusion. The fluorophore Cy5 was covalently coupled to digoxigenin and quantitatively bound by the bispecific antibody. A panel of tumor cell lines was assessed under different culture conditions for absolute receptor expression levels of the indicated antigens and the data were set in relation to mRNA, gene count and immunoblot data. We could reproducibly quantify these receptors, omit the otherwise required normalization step and demonstrate the superiority of a 1 + 1 bispecific antibody. The same antibodies were also used to quantify the number of proteins in intracellular vesicles in confocal microscopy. The antibodies can be stored like regular antibodies and can be coupled with different digoxigenin-labeled fluorophores which makes them excellent tools for FACS and imaging-based experiments. PMID:23960142

Panke, C; Weininger, D; Haas, A; Schelter, F; Schlothauer, T; Bader, S; Sircar, R; Josel, H P; Baer, U; Burtscher, H; Mundigl, O; Grote, M; Brinkmann, U; Sustmann, C

2013-10-01

34

Bispecific small molecule-antibody conjugate targeting prostate cancer  

PubMed Central

Bispecific antibodies, which simultaneously target CD3 on T cells and tumor-associated antigens to recruit cytotoxic T cells to cancer cells, are a promising new approach to the treatment of hormone-refractory prostate cancer. Here we report a site-specific, semisynthetic method for the production of bispecific antibody-like therapeutics in which a derivative of the prostate-specific membrane antigen-binding small molecule DUPA was selectively conjugated to a mutant ?CD3 Fab containing the unnatural amino acid, p-acetylphenylalanine, at a defined site. Homogeneous conjugates were generated in excellent yields and had good solubility. The efficacy of the conjugate was optimized by modifying the linker structure, relative binding orientation, and stoichiometry of the ligand. The optimized conjugate showed potent and selective in vitro activity (EC50 ?100 pM), good serum half-life, and potent in vivo activity in prophylactic and treatment xenograft mouse models. This semisynthetic approach is likely to be applicable to the generation of additional bispecific agents using drug-like ligands selective for other cell-surface receptors.

Kim, Chan Hyuk; Axup, Jun Y.; Lawson, Brian R.; Yun, Hwayoung; Tardif, Virginie; Choi, Sei Hyun; Zhou, Quan; Dubrovska, Anna; Biroc, Sandra L.; Marsden, Robin; Pinstaff, Jason; Smider, Vaughn V.; Schultz, Peter G.

2013-01-01

35

Bispecific small molecule-antibody conjugate targeting prostate cancer.  

PubMed

Bispecific antibodies, which simultaneously target CD3 on T cells and tumor-associated antigens to recruit cytotoxic T cells to cancer cells, are a promising new approach to the treatment of hormone-refractory prostate cancer. Here we report a site-specific, semisynthetic method for the production of bispecific antibody-like therapeutics in which a derivative of the prostate-specific membrane antigen-binding small molecule DUPA was selectively conjugated to a mutant ?CD3 Fab containing the unnatural amino acid, p-acetylphenylalanine, at a defined site. Homogeneous conjugates were generated in excellent yields and had good solubility. The efficacy of the conjugate was optimized by modifying the linker structure, relative binding orientation, and stoichiometry of the ligand. The optimized conjugate showed potent and selective in vitro activity (EC50 ~ 100 pM), good serum half-life, and potent in vivo activity in prophylactic and treatment xenograft mouse models. This semisynthetic approach is likely to be applicable to the generation of additional bispecific agents using drug-like ligands selective for other cell-surface receptors. PMID:24127589

Kim, Chan Hyuk; Axup, Jun Y; Lawson, Brian R; Yun, Hwayoung; Tardif, Virginie; Choi, Sei Hyun; Zhou, Quan; Dubrovska, Anna; Biroc, Sandra L; Marsden, Robin; Pinstaff, Jason; Smider, Vaughn V; Schultz, Peter G

2013-10-29

36

Cell-Penetrating Bispecific Antibodies for Targeting Oncogenic Transcription Factors in Advanced Prostate Cancer.  

National Technical Information Service (NTIS)

The goal of these studies is to develop and validate cell- penetrating bi-specific antibodies as an agent that can selectively inhibit the function of intracellular proteins. We have developed 3E10-AR441 bi-specific antibody to inhibit the function both l...

M. Lilly

2013-01-01

37

Bispecific Antibody Pretargeting for Improving Cancer Imaging and Therapy  

SciTech Connect

The main objective of this project was to evaluate pretargeting systems that use a bispecific antibody (bsMAb) to improve the detection and treatment of cancer. A bsMAb has specificity to a tumor antigen, which is used to bind the tumor, while the other specificity is to a peptide that can be radiolabeled. Pretargeting is the process by which the unlabeled bsMAb is given first, and after a sufficient time (1-2 days) is given for it to localize in the tumor and clear from the blood, a small molecular weight radiolabeled peptide is given. According to a dynamic imaging study using a 99mTc-labeled peptide, the radiolabeled peptide localizes in the tumor in less than 1 hour, with > 80% of it clearing from the blood and body within this same time. Tumor/nontumor targeting ratios that are nearly 50 times better than that with a directly radiolabeled Fab fragment have been observed (Sharkey et al., ''Signal amplification in molecular imaging by a multivalent bispecific nanobody'' submitted). The bsMAbs used in this project have been composed of 3 antibodies that will target antigens found in colorectal and pancreatic cancers (CEA, CSAp, and MUC1). For the ''peptide binding moiety'' of the bsMAb, we initially examined an antibody directed to DOTA, but subsequently focused on another antibody directed against a novel compound, HSG (histamine-succinyl-glycine).

Sharkey, Robert M.

2005-02-04

38

Monoclonal antibodies in oncology.  

PubMed Central

Molecular biology has made tremendous strides over the last five years. The new biology allows us to prepare monoclonal antibodies to defined antigens; to detect, isolate and clone specific genes; and to insert these genes into defined sites in different cells giving new functions to old organisms. These revolutionary developments have been followed closely by researchers, businessmen, politicians and philosophers, as well as by those involved in the clinical care of patients. Although our understanding of human molecular biology is increasing rapidly, it is the development of monoclonal antibodies that has the most immediate application in the clinic. There have been several reports of their use in the diagnosis, localisation and treatment of human malignant disease. This review describes developments that are likely to have direct relevance to patient care in the near future.

Sikora, K

1982-01-01

39

Ipilimumab augments antitumor activity of bispecific antibody-armed T cells  

PubMed Central

Background Ipilimumab is an antagonistic monoclonal antibody against cytotoxic T-lymphocyte antigen-4 (CTLA-4) that enhances antitumor immunity by inhibiting immunosuppressive activity of regulatory T cells (Treg). In this study, we investigated whether inhibiting Treg activity with ipilimumab during ex vivo T cell expansion could augment anti-CD3-driven T cell proliferation and enhance bispecific antibody (BiAb)-redirected antitumor cytotoxicity of activated T cells (ATC). Methods PBMC from healthy individuals were stimulated with anti-CD3 monoclonal antibody with or without ipilimumab and expanded for 10-14 days. ATC were harvested and armed with anti-CD3 x anti-EGFR BiAb (EGFRBi) or anti-CD3 x anti-CD20 BiAb (CD20Bi) to test for redirected cytotoxicity against COLO356/FG pancreatic cancer cell line or Burkitt’s lymphoma cell line (Daudi). Results In PBMC from healthy individuals, the addition of ipilimumab at the initiation of culture significantly enhanced T cell proliferation (p?=?0.0029). ATC grown in the presence of ipilimumab showed significantly increased mean tumor-specific cytotoxicity at effector:target (E:T) ratio of 25:1 directed at COLO356/FG and Daudi by 37.71% (p?bispecific antibodies.

2014-01-01

40

Monospecific and bispecific antibodies against E. coli O157 for diagnostics.  

PubMed

Escherichia coli O157:H7 is a serious human pathogen that causes hemorrhagic colitis, and occasionally hemolytic uremic syndrome. Identification of the O157 antigen is an essential part of the detection and management of E. coli O157:H7. A quadroma P126 secreting a bispecific hybrid MAb (bsMAb), which recognizes both E. coli O157 and horseradish peroxidase in one molecule was produced by somatic hybridization of hybridomas specific for E. coli O157 and HRPO molecule. A bridge ELISA was used to select the quadromas obtained for bispecific monoclonal antibody purification and characterization. Benzhydroxamic-acid agarose (BHA) affinity co-chromatography was used as a convenient one-step method for purifying the HRPO-bsMAb complex for ultrasensitive diagnostic applications. Sandwich ELISA for detecting E. coli O157:H7 with HRPO-bsMAb allows quick one step detection of spiked E. coli O157:H7. The detection sensitivities were 100 CFU, 750 CFU and 500 CFU per 1 ml of tap water, lake water and apple juice respectively by microtiter assay. E. coli O157:H7 detection with immunofilter ELISA and immunomagnetic ELISA formats was approximately 1 CFU/ml and 10 CFU/ml respectively. BsMAbs avoid enzyme conjugation, has highest specific activity and molecular uniformity without aggregates and contribute to good signal to noise ratios. This new bispecific antibody can be generated and purified from quadroma cultures by affinity co-chromatography in one step and can be used to develop a new generation of assays for public health applications in water, food and human sample testing. PMID:17804009

Guttikonda, Sujatha; Tang, Xinling L; Yang, Bozka M; Armstrong, Glen D; Suresh, Mavanur R

2007-10-31

41

Biodistribution studies with tumor-targeting bispecific antibodies reveal selective accumulation at the tumor site.  

PubMed

Bispecific antibodies are proteins that bind two different antigens and may retarget immune cells with a binding moiety specific for a leukocyte marker. A binding event in blood could in principle prevent antibody extravasation and accumulation at the site of disease. In this study, we produced and characterized two tetravalent bispecific antibodies that bind with high affinity to the alternatively-spliced EDB domain of fibronectin, a tumor-associated antigen. The bispecific antibodies simultaneously engaged the cognate antigens (murine T cell co-receptor CD3 and hen egg lysozyme) and selectively accumulated on murine tumors in vivo. The results, which were in agreement with predictions based on pharmacokinetic modeling and antibody binding characteristics, confirmed that bispecific antibodies can reach abluminal targets without being blocked by peripheral blood leukocytes. PMID:23032949

List, Thomas; Neri, Dario

2012-01-01

42

Norovirus Monoclonal Antibodies and Peptides.  

National Technical Information Service (NTIS)

The present invention is drawn to monoclonal antibodies that bind to a Norovirus, peptides that inhibit monoclonal antibody binding to a Norovirus, and peptides that inhibit binding of a Norovirus to a cell. The compositions of the invention find use as N...

M. Hardy

2004-01-01

43

Chemically Programmed Bispecific Antibodies That Recruit and Activate T Cells*  

PubMed Central

Bispecific antibodies (biAbs) that mediate cytotoxicity by recruiting and activating endogenous immune cells are an emerging class of next-generation antibody therapeutics. Of particular interest are biAbs of relatively small size (?50 kDa) that can redirect cytotoxic T cells through simultaneous binding of tumor cells. Here we describe a conceptually unique class of biAbs in which the tumor cell specificity of a humanized antibody fragment that recognizes CD3 on T cells is chemically programmed through a C-terminal selenocysteine (Sec) residue. We demonstrate that through chemically programmed specificity for integrin ?4?1 or folate receptor 1 (FOLR1), and common specificity for CD3, these hybrid molecules exert potent and specific in vitro and ex vivo cytotoxicity toward tumor cell lines and primary tumor cells in the presence of primary T cells. Importantly, the generic nature of chemical programming allows one to apply our approach to virtually any specificity, promising a broad utility of chemically programmed biAbs in cancer therapy.

Cui, Huiting; Thomas, Joshua D.; Burke, Terrence R.; Rader, Christoph

2012-01-01

44

Chemically programmed bispecific antibodies that recruit and activate T cells.  

PubMed

Bispecific antibodies (biAbs) that mediate cytotoxicity by recruiting and activating endogenous immune cells are an emerging class of next-generation antibody therapeutics. Of particular interest are biAbs of relatively small size (?50 kDa) that can redirect cytotoxic T cells through simultaneous binding of tumor cells. Here we describe a conceptually unique class of biAbs in which the tumor cell specificity of a humanized antibody fragment that recognizes CD3 on T cells is chemically programmed through a C-terminal selenocysteine (Sec) residue. We demonstrate that through chemically programmed specificity for integrin ?(4)?(1) or folate receptor 1 (FOLR1), and common specificity for CD3, these hybrid molecules exert potent and specific in vitro and ex vivo cytotoxicity toward tumor cell lines and primary tumor cells in the presence of primary T cells. Importantly, the generic nature of chemical programming allows one to apply our approach to virtually any specificity, promising a broad utility of chemically programmed biAbs in cancer therapy. PMID:22761439

Cui, Huiting; Thomas, Joshua D; Burke, Terrence R; Rader, Christoph

2012-08-17

45

Monoclonal Antibody Pharmacokinetics and Pharmacodynamics  

Microsoft Academic Search

More than 20 monoclonal antibodies have been approved as therapeutic drugs by the US Food and Drug Administration, and it is quite likely that the number of approved antibodies will double in the next 7–10 years. Antibody drugs show several desirable characteristics, including good solubility and stability, long persistence in the body, high selectivity and specificity, and low risk for

W Wang; EQ Wang; JP Balthasar

2008-01-01

46

Generating bispecific human IgG1 and IgG2 antibodies from any antibody pair.  

PubMed

Bispecific antibodies and antibody fragments are a new class of therapeutics increasingly utilized in the clinic for T cell recruitment (catumaxomab anti-EpCAM/CD3 and blinatumomab anti-CD19/CD3), increase in the selectivity of targeting, or simultaneous modulation of multiple cellular pathways. While the clinical potential for certain bispecific antibody formats is clear, progress has been hindered because they are often difficult to manufacture, may suffer from suboptimal pharmacokinetic properties, and may be limited due to potential immunogenicity issues. Current state-of-the-art human IgG-like bispecific technologies require co-expression of two heavy chains with a single light chain, use crossover domains to segregate light chains, or utilize scFv (single-chain fragment variable)-Fc fusion. We have engineered both human IgG1 and IgG2 subtypes, with minimal point mutations, to form full-length bispecific human antibodies with high efficiency and in high purity. In our system, the two antibodies of interest can be expressed and purified separately, mixed together under appropriate redox conditions, resulting in a formation of a stable bispecific antibody with high yields. With this approach, it is not necessary to generate new antibodies that share a common light chain, therefore allowing the immediate use of an existing antibody regardless of whether it has been generated via standard hybridoma or display methods. We demonstrate the generality of the approach and show that these bispecific antibodies have properties similar to those of wild-type IgGs, and we further demonstrate the utility of the technology with an example of a CD3/CD20 bispecific antibody that effectively depletes B cells in vitro and in vivo. PMID:22543237

Strop, Pavel; Ho, Wei-Hsien; Boustany, Leila M; Abdiche, Yasmina N; Lindquist, Kevin C; Farias, Santiago E; Rickert, Mathias; Appah, Charles Takeshi; Pascua, Edward; Radcliffe, Teresa; Sutton, Janette; Chaparro-Riggers, Javier; Chen, Wei; Casas, Meritxell Galindo; Chin, Sherman Michael; Wong, Oi Kwan; Liu, Shu-Hui; Vergara, German; Shelton, Dave; Rajpal, Arvind; Pons, Jaume

2012-07-13

47

Immunotoxicity of monoclonal antibodies  

PubMed Central

Monoclonal antibodies (mAbs) are large molecules intended to bind to specific targets often expressed on the immune system, and to treat various immunopathological conditions. Therefore, mAbs can be considered to have a high potential for immunotoxicity, which is reflected in the clinical experience accumulated on mAbs-induced adverse effects related to immunosuppression, immunostimulation and hypersensitivity (immunogenicity). So far, non clinical immunotoxicity studies have been inadequate to address all safety issues in relation to the possible immunotoxicity of mAbs, because they are fraught with limitations and pitfalls primarily related to the lack of relevant animal species. In addition, clinical studies rarely include validated end-points dedicated to the prediction of immunotoxicity. With the ongoing development of mAbs as novel therapeutic strategies for a wide variety of diseases, efforts should be paid to improve our understanding of mAbs-induced immunotoxic effects and design dedicated strategies to assess their immunological safety, both non clinically and clinically.

2009-01-01

48

Rational design and generation of recombinant control reagents for bispecific antibodies through CDR mutagenesis.  

PubMed

Developments in the field of bispecific antibodies have progressed rapidly in recent years, particularly in their potential role for the treatment of malignant disease. However, manufacturing stable molecules has proven to be costly and time-consuming, which in turn has hampered certain aspects of preclinical evaluation including the unavailability of appropriate "negative" controls. Bispecific molecules (e.g., bispecific tandem scFv) exhibit two specificities, often against a tumor antigen as well as an immune-activation ligand such as CD3. While for IgG antibodies, isotype-matched controls are well accepted, when considering smaller antibody fragments it is not possible to adequately control for their biological activity through the use of archetypal isotypes, which differ dramatically in affinity, size, structure, and design. Here, we demonstrate a method for the rapid production of negative control tandem scFvs through complementarity determining region (CDR) mutagenesis, using a recently described bispecific T-cell engager (BiTE) targeting a tumor-specific mutation of the epidermal growth factor receptor (EGFRvIII) as an example. Four independent control constructs were developed by this method through alteration of residues spanning individual CDR domains. Importantly, while target antigen affinity was completely impaired, CD3 binding affinity was conserved in each molecule. These results have a potential to enhance the sophistication by which bispecific antibodies can be evaluated in the preclinical setting and may have broader applications for an array of alternative antibody-derived therapeutic platforms. PMID:23806556

Choi, Bryan D; Gedeon, Patrick C; Kuan, Chien-Tsun; Sanchez-Perez, Luis; Archer, Gary E; Bigner, Darell D; Sampson, John H

2013-09-30

49

Structural understanding of stabilization patterns in engineered bispecific Ig-like antibody molecules  

SciTech Connect

Bispecific immunoglobulin-like antibodies capable of engaging multiple antigens represent a promising new class of therapeutic agents. Engineering of these molecules requires optimization of the molecular properties of one of the domain components. Here, we present a detailed crystallographic and computational characterization of the stabilization patterns in the lymphotoxin-beta receptor (LT{beta}R) binding Fv domain of an anti-LT{beta}R/anti-TNF-related apoptosis inducing ligand receptor-2 (TRAIL-R2) bispecific immunoglobulin-like antibody. We further describe a new hierarchical structure-guided approach toward engineering of antibody-like molecules to enhance their thermal and chemical stability.

Jordan, Jacob L.; Arndt, Joseph W.; Hanf, Karl; Li, Guohui; Hall, Janine; Demarest, Stephen; Huang, Flora; Wu, Xiufeng; Miller, Brian; Glaser, Scott; Fernandez, Erik J.; Wang, Deping; Lugovskoy, Alexey; (UV); (Biogen)

2010-01-12

50

Heavy chain-only antibodies and tetravalent bispecific antibody neutralizing Staphylococcus aureus leukotoxins.  

PubMed

Panton-Valentine leukocidin (PVL) is a pore-forming toxin associated with current outbreaks of community-associated methicillin-resistant strains and implicated directly in the pathophysiology of Staphylococcus aureus-related diseases. Humanized heavy chain-only antibodies (HCAb) were generated against S. aureus PVL from immunized transgenic mice to neutralize toxin activity. The active form of PVL consists of the two components, LukS-PV and LukF-PV, which induce osmotic lysis following pore formation in host defense cells. One anti-LukS-PV HCAb, three anti-LukF-PV HCAbs with affinities in the nanomolar range, and one engineered tetravalent bispecific HCAb were tested in vitro and in vivo, and all prevented toxin binding and pore formation. Anti-LukS-PV HCAb also binds to ?-hemolysin C (HlgC) and inhibits HlgC/HlgB pore formation. Experiments in vivo in a toxin-induced rabbit endophthalmitis model showed that these HCAbs inhibit inflammatory reactions and tissue destruction, with the tetravalent bispecific HCAb performing best. Our findings show the therapeutic potential of HCAbs, and in particular, bispecific antibodies. PMID:21930905

Laventie, Benoît-Joseph; Rademaker, Hendrik Jan; Saleh, Maher; de Boer, Ernie; Janssens, Rick; Bourcier, Tristan; Subilia, Audrey; Marcellin, Luc; van Haperen, Rien; Lebbink, Joyce H G; Chen, Tao; Prévost, Gilles; Grosveld, Frank; Drabek, Dubravka

2011-09-27

51

Fragmentation of monoclonal antibodies.  

PubMed

Fragmentation is a degradation pathway ubiquitously observed in proteins despite the remarkable stability of peptide bond; proteins differ only by how much and where cleavage occurs. The goal of this review is to summarize reports regarding the non-enzymatic fragmentation of the peptide backbone of monoclonal antibodies (mAbs). The sites in the polypeptide chain susceptible to fragmentation are determined by a multitude of factors. Insights are provided on the intimate chemical mechanisms that can make some bonds prone to cleavage due to the presence of specific side-chains. In addition to primary structure, the secondary, tertiary and quaternary structures have a significant impact in modulating the distribution of cleavage sites by altering local flexibility, accessibility to solvent or bringing in close proximity side chains that are remote in sequence. This review focuses on cleavage sites observed in the constant regions of mAbs, with special emphasis on hinge fragmentation. The mechanisms responsible for backbone cleavage are strongly dependent on pH and can be catalyzed by metals or radicals. The distribution of cleavage sites are different under acidic compared to basic conditions, with fragmentation rates exhibiting a minimum in the pH range 5 to 6; therefore, the overall fragmentation pattern observed for a mAb is a complex result of structural and solvent conditions. A critical review of the techniques used to monitor fragmentation is also presented; usually a compromise has to be made between a highly sensitive method with good fragment separation and the capability to identify the cleavage site. The effect of fragmentation on the function of a mAb must be evaluated on a case-by-case basis depending on whether cleavage sites are observed in the variable or constant regions, and on the mechanism of action of the molecule. PMID:21487244

Vlasak, Josef; Ionescu, Roxana

2011-01-01

52

Unexpected recombinations in single chain bispecific anti-CD3-anti-CD33 antibodies can be avoided by a novel linker module.  

PubMed

CD33 is an attractive immunotarget on the surface of tumor cells from patients with acute myeloid leukemia (AML). In a first attempt for immunotargeting of AML blasts we constructed two bispecific antibodies in the single chain bispecific diabody (scBsDb) format by fusing the variable domains of monoclonal antibodies directed against CD3 and CD33. Unfortunately, protein expression of both scBsDbs resulted in varying mixtures of fragmented and full length proteins. As the non-functional fragments competed with the functional full length antibodies we tried to understand the reason for the fragmentation. We found that the anti-CD3 and anti-CD33 antibody genes show striking sequence homologies: during B cell development the same V(h) J558 heavy and V(l) kk4 light chain genes were selected. Moreover, the closely related D genes DSP2 (9 and 11) were combined with the same JH4 gene. And finally, during VJ recombination of the light chain the same JK5 element was selected. These homologies between the two monoclonal antibodies were the reason for recombinations in the cell lines generated for expression of the scBsDbs. Finally, we solved this problem by (i) rearranging the order of the heavy and light chains of the anti-CD3 and anti-CD33 domains, and (ii) a replacement of one of the commonly used glycine serine linkers with a novel linker domain. The resulting bispecific antibody in a single chain bispecific tandem format (scBsTaFv) was stable and capable of redirecting T cells to CD33-positive tumor cells including AML blasts of patients. PMID:22014687

Stamova, Slava; Cartellieri, Marc; Feldmann, Anja; Arndt, Claudia; Koristka, Stefanie; Bartsch, Holger; Bippes, Claudia C; Wehner, Rebekka; Schmitz, Marc; von Bonin, Malte; Bornhäuser, Martin; Ehninger, Gerhard; Bachmann, Michael

2011-12-01

53

Cooperative mixtures of bispecific F(ab')2 antibodies for delivering saporin to lymphoma in vitro and in vivo  

SciTech Connect

We report that selected combinations of two or more monoclonal bispecific F(ab')2 antibodies (BsAbs) far outperform single derivatives in the delivery of the ribosome-inactivating protein, saporin, to guinea pig L2C leukemic cells. Throughout the work, BsAbs were constructed by thioether-linking the hinges of two Fab'gamma, one from monoclonal anti-L2C-idiotype antibody and the other from anti-saporin antibody. The latter was either affinity-purified rabbit polyclonal or one of a panel of five mouse monoclonal antibodies. In vitro cytotoxicity studies showed that, though all derivatives were effective, the BsAb made with the polyclonal antibody was always 10 to 20 times more potent than those made with a monoclonal antibody in yielding 50% inhibition of (3H)leucine uptake. This superior activity could be matched by selective mixtures of two or more of the monoclonal derivatives. Furthermore, in immunotherapeutic delivery of saporin to tumor, a pair of BsAbs performed significantly better than did either individually. Binding and uptake studies with radiolabeled saporin demonstrated a 20-fold increase in functional affinity when saporin was held at the cell surface by an appropriate BsAb mixture rather than by a single BsAb. In contrast, only small differences were recorded in the rate at which saporin was internalized as a result of the same maneuver. We conclude that the improved performance of combinations of BsAbs arises from their ability to provide multiple linkages between saporin molecules and cell surfaces, increasing the functional affinity with which saporin is tethered to the cell, but, in this system at least, having only a minor effect on the rate at which it is internalized. Cocktails of two or more BsAbs, selected to bind to multiple epitopes on ribosome-inactivating proteins could provide an important new strategy in immunotherapy.

French, R.R.; Courtenay, A.E.; Ingamells, S.; Stevenson, G.T.; Glennie, M.J. (General Hospital, Southampton (England))

1991-05-01

54

Bispecific antibodies and trispecific immunocytokines for targeting the immune system against cancer: preparing for the future.  

PubMed

Monoclonal anti-tumor antibodies (mAbs) that are clinically effective usually recruit, via their constant fragment (Fc) domain, Fc receptor (FcR)-positive accessory cells of the immune system and engage these additionally against the tumor. Since T cells are FcR negative, these important cells are not getting involved. In contrast to mAbs, bispecific antibodies (bsAbs) can be designed in such a way that they involve T cells. bsAbs are artificially designed molecules that bind simultaneously to two different antigens, one on the tumor cell, the other one on an immune effector cell such as CD3 on T cells. Such dual antibody constructs can cross-link tumor cells and T cells. Many such bsAb molecules at the surface of tumor cells can thus build a bridge to T cells and aggregate their CD3 molecules, thereby activating them for cytotoxic activity. BsAbs can also contain a third binding site, for instance a Fc domain or a cytokine that would bind to its respective cytokine receptor. The present review discusses the pros and cons for the use of the Fc fragment during the development of bsAbs using either cell-fusion or recombinant DNA technologies. The recombinant antibody technology allows the generation of very efficient bsAbs containing no Fc domain such as the bi-specific T-cell engager (BiTE). The strong antitumor activity of these molecules makes them very interesting new cancer therapeutics. Over the last decade, we have developed another concept, namely to combine bsAbs and multivalent immunocytokines with a tumor cell vaccine. The latter are patient-derived tumor cells modified by infection with a virus. The virus-Newcastle Disease Virus (NDV)-introduces, at the surface of the tumor cells, viral molecules that can serve as general anchors for the bsAbs. Our strategy aims at redirecting, in an Fc-independent fashion, activities of T cells and accessory cells against autologous tumor antigens. It creates very promising perspectives for a new generation of efficient and safe cancer therapeutics that should confer long-lasting anti-tumor immunity. PMID:23329400

Fournier, Philippe; Schirrmacher, Volker

2013-02-01

55

Uses of monoclonal antibody 8H9  

DOEpatents

This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a pharmaceutically acceptable carrier. This invention also provides an antibody other than the monoclonal antibody 8H9 comprising the complementary determining regions of monoclonal antibody 8H9 or a derivative thereof, capable of binding to the same antigen as the monoclonal antibody 8H9. This invention provides a substance capable of competitively inhibiting the binding of monoclonal antibody 8H9. This invention also provides an isolated scFv of monoclonal antibody 8H9 or a derivative thereof. This invention also provides the 8H9 antigen. This invention also provides different uses of the monoclonal antibody 8H9 or its derivative.

Cheung, Nai-Kong V.

2013-04-09

56

BiTEs: bispecific antibody constructs with unique anti-tumor activity.  

PubMed

Bispecific T-cell engager molecules (BiTEs) constitute a class of bispecific single-chain antibodies for the polyclonal activation and redirection of cytotoxic T cells against pathogenic target cells. BiTEs combine a unique set of properties that have not yet been reported for any other kind of bispecific antibody construct, namely extraordinary potency and efficacy against target cells at low T-cell numbers without the need for T-cell co-stimulation. Here we review novel insights into the mechanism of BiTE action, which help to explain the unique features of BiTEs, as well as data from various animal models demonstrating the outstanding therapeutic potential of BiTEs for the treatment of malignant diseases. PMID:16213416

Wolf, Evelyn; Hofmeister, Robert; Kufer, Peter; Schlereth, Bernd; Baeuerle, Patrick A

2005-09-15

57

Monoclonal antibodies for treating cancer  

SciTech Connect

The purpose of this study is to assess the current status of in-vivo use of monoclonal antibodies for treating cancer. Publications appearing between 1980 and 1988 were identified by computer searches using MEDLINE and CANCERLIT, by reviewing the table of contents of recently published journals, and by searching bibliographies of identified books and articles. More than 700 articles, including peer-reviewed articles and book chapters, were identified and selected for analysis. The literature was reviewed and 235 articles were selected as relevant and representative of the current issues and future applications for in-vivo monoclonal antibodies for cancer therapy and of the toxicity and efficacy which has been associated with clinical trials. Approaches include using antibody alone (interacting with complement or effector cells or binding directly with certain cell receptors) and immunoconjugates (antibody coupled to radioisotopes, drugs, toxins, or other biologicals). Most experience has been with murine antibodies. Trials of antibody alone and radiolabeled antibodies have confirmed the feasibility of this approach and the in-vivo trafficking of antibodies to tumor cells. However, tumor cell heterogeneity, lack of cytotoxicity, and the development of human antimouse antibodies have limited clinical efficacy. Although the immunoconjugates are very promising, heterogeneity and the antimouse immune response have hampered this approach as has the additional challenge of chemically or genetically coupling antibody to cytotoxic agents. As a therapeutic modality, monoclonal antibodies are still promising but their general use will be delayed for several years. New approaches using human antibodies and reducing the human antiglobulin response should facilitate treatment. 235 references.

Dillman, R.O. (Hoag Cancer Center, Newport Beach, CA (USA))

1989-10-01

58

Detection of Campylobacter species using monoclonal antibodies  

NASA Astrophysics Data System (ADS)

A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

Young, Colin R.; Lee, Alice; Stanker, Larry H.

1999-01-01

59

Uses of Monoclonal Antibody 8H9.  

National Technical Information Service (NTIS)

This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 ...

N. K. Cheung

2003-01-01

60

Platelet Fibrinogen-Specific Monoclonal Antibody.  

National Technical Information Service (NTIS)

The invention is related generally to monoclonal antibodies. More particularly, the invention is related to a unique monoclonal antibody (MAb) directed against a specific antigen, the platelet fibrinogen, found on stimulated platelets, said MAb being desi...

H. Grelnick

1990-01-01

61

Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies  

PubMed Central

The development of bispecific antibodies has attracted substantial interest, and many different formats have been described. Those specifically containing an Fc part are mostly tetravalent, such as stabilized IgG-scFv fusions or dual-variable domain (DVD) IgGs. However, although they exhibit IgG-like properties and technical developability, these formats differ in size and geometry from classical IgG antibodies. Thus, considerable efforts focus on bispecific heterodimeric IgG antibodies that more closely mimic natural IgG molecules. The inherent chain association problem encountered when producing bispecific heterodimeric IgG antibodies can be overcome by several methods. While technologies like knobs-into-holes (KiH) combined with a common light chain or the CrossMab technology enforce the correct chain association, other approaches, e.g., the dual-acting Fab (DAF) IgGs, do not rely on a heterodimeric Fc part. This review discusses the state of the art in bispecific heterodimeric IgG antibodies, with an emphasis on recent progress.

Klein, Christian; Sustmann, Claudio; Thomas, Markus; Stubenrauch, Kay; Croasdale, Rebecca; Schanzer, Jurgen; Brinkmann, Ulrich; Kettenberger, Hubert; Regula, Jorg T.; Schaefer, Wolfgang

2012-01-01

62

Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies.  

PubMed

The development of bispecific antibodies has attracted substantial interest, and many different formats have been described. Those specifically containing an Fc part are mostly tetravalent, such as stabilized IgG-scFv fusions or dual-variable domain (DVD) IgGs. However, although they exhibit IgG-like properties and technical developability, these formats differ in size and geometry from classical IgG antibodies. Thus, considerable efforts focus on bispecific heterodimeric IgG antibodies that more closely mimic natural IgG molecules. The inherent chain association problem encountered when producing bispecific heterodimeric IgG antibodies can be overcome by several methods. While technologies like knobs-into-holes (KiH) combined with a common light chain or the CrossMab technology enforce the correct chain association, other approaches, e.g., the dual-acting Fab (DAF) IgGs, do not rely on a heterodimeric Fc part. This review discusses the state of the art in bispecific heterodimeric IgG antibodies, with an emphasis on recent progress. PMID:22925968

Klein, Christian; Sustmann, Claudio; Thomas, Markus; Stubenrauch, Kay; Croasdale, Rebecca; Schanzer, Jürgen; Brinkmann, Ulrich; Kettenberger, Hubert; Regula, Jörg T; Schaefer, Wolfgang

2012-01-01

63

Engineering Bispecific Antibodies that Target ErbB-2 on Breast Cancer Cells.  

National Technical Information Service (NTIS)

The purpose of this project is twofold. First, to construct novel bispecific antibodies that can be expressed as a single-chain in E. coli.. Second, to develop an in vivo animal model, using a TCR transgenic/RAG(-/-) strain of mice, for evaluating agents ...

D. M. Kranz

1997-01-01

64

Application of bispecific antibody against antigen and hapten for immunodetection and immunopurification  

PubMed Central

We present a bispecific antibody that recognizes an antigen and a hapten and can be applied to various biological assays, including immunoblotting and immunoprecipitation. In immunoblot analysis of serum, an anti-C5 × anti-cotinine bispecific tandem single-chain variable fragment (scFv)-Fc fusion protein and cotinine-conjugated horseradish peroxidase (HRP) generated a clean signal without the high background that was observed in a parallel experiment using HRP-conjugated goat anti-rabbit immunoglobulin G (Fc-specific) antibody. In immunoprecipitation analysis of serum, use of the bispecific tandem scFv-Fc fusion protein and cotinine-crosslinked magnetic beads significantly reduced the amount of protein contaminants compared with a parallel experiment done with protein A agarose beads. In subsequent immunoblot analysis, use of cotinine–HRP as the secondary probe instead of HRP-conjugated goat anti-rabbit IgG (Fc-specific) antibody successfully eliminated the band corresponding to the bispecific tandem scFv-Fc fusion protein.

Kim, Hyori; Park, Sunyoung; Lee, Hwa Kyoung; Chung, Junho

2013-01-01

65

Monoclonal antibodies in haematopathology  

SciTech Connect

This book contains over 40 selections. Some of the titles are: Oncogene (c-myc, c-myb) amplification in acute myelogenous leukaemia; Ultrastructural characterization of leukaemic cells with monoloclonal antibodies; Origin of B-cell malignancies; Immunohistology of gut lymphomas; and Spurious evidence of lineage infidelity in monocytic leukaemia.

Grignani, F.; Martelli, M.F.; Mason, D.Y.

1985-01-01

66

Generation of single-chain bispecific green fluorescent protein fusion antibodies for imaging of antibody-induced T cell synapses.  

PubMed

There is growing interest in the development of novel single-chain bispecific antibodies for retargeting of immune effector T cells to tumor cells. Until today, functional fusion constructs consisting of a single-chain bispecific antibody and a fluorescent protein were not reported. Such molecules could be useful for an in vivo visualization of this retargeting process. Recently, we established two novel single-chain bispecific antibodies. One is capable of retargeting T cells to CD33, and the other is capable of retargeting T cells to the prostate stem cell antigen (PSCA). CD33 is an attractive immunotarget on the surface of tumor cells from patients with acute myeloid leukemia (AML). The PSCA is a potential target on prostate cancer cells. Flanking the reading frame encoding the green fluorescent protein (GFP) with a recently described novel helical linker element allowed us to establish novel single-chain bispecific fusion antibodies. These fluorescent fusion antibodies were useful to efficiently retarget T cells to the respective tumor cells and visualize the formation of immune synapses between effector and target cells. PMID:22274538

Stamova, Slava; Feldmann, Anja; Cartellieri, Marc; Arndt, Claudia; Koristka, Stefanie; Apel, Falko; Wehner, Rebekka; Schmitz, Marc; Bornhäuser, Martin; von Bonin, Malte; Ehninger, Gerhard; Bartsch, Holger; Bachmann, Michael

2012-04-15

67

Radioimmunoguided surgery using monoclonal antibody  

SciTech Connect

The potential proficiency of radioimmunoguided surgery in the intraoperative detection of tumors was assessed using labeled monoclonal antibody B72.3 in 66 patients with tissue-proved tumor. Monoclonal antibody B72.3 was injected 5 to 42 days preoperatively, and the hand-held gamma-detecting probe was used intraoperatively to detect the presence of tumor. Intraoperative probe counts of less than 20 every 2 seconds, or tumor-to-adjacent normal tissue ratios less than 2:1 were considered negative (system failure). Positive probe counts were detected in 5 of 6 patients with primary colon cancer (83 percent), in 31 of 39 patients with recurrent colon cancer (79 percent), in 4 of 5 patients with gastric cancer (80 percent), in 3 of 8 patients with breast cancer (37.5 percent), and in 4 of 8 patients with ovarian cancer (50 percent) undergoing second-look procedures. Additional patients in each group were scored as borderline positive. Overall, radioimmunoguided surgery using B72.3 identified tumors in 47 patients (71.2 percent), bordered on positive in 6 patients (9.1 percent), and failed to identify tumor in 13 patients (19.7 percent). Improved selection of patients for antigen-positive tumors, the use of higher affinity second-generation antibodies, alternate routes of antibody administration, alternate radionuclides, and more sophisticatedly bioengineered antibodies and antibody combinations should all lead to improvements in radioimmunoguided surgery.

Martin, E.W. Jr.; Mojzisik, C.M.; Hinkle, G.H. Jr.; Sampsel, J.; Siddiqi, M.A.; Tuttle, S.E.; Sickle-Santanello, B.; Colcher, D.; Thurston, M.O.; Bell, J.G.

1988-11-01

68

A novel bispecific antibody format enables simultaneous bivalent and monovalent co-engagement of distinct target antigens.  

PubMed

Bispecific antibodies based on full-length antibody structures are more optimal than fragment-based formats because they benefit from the favorable properties of the Fc region. However, the homodimeric nature of Fc effectively imposes bivalent binding on all current full-length bispecific antibodies, an attribute that can result in nonspecific activation of cross-linked receptors. We engineered a novel bispecific format, referred to as mAb-Fv, that utilizes a heterodimeric Fc region to enable monovalent co-engagement of a second target antigen in a full-length context. mAb-Fv constructs co-targeting CD16 and CD3 were expressed and purified as heterodimeric species, bound selectively to their co-target antigens, and mediated potent cytotoxic activity by NK cells and T cells, respectively. The capacity to co-engage distinct target antigens simultaneously with different valencies is an improved feature for bispecific antibodies with promising therapeutic implications. PMID:22123055

Moore, Gregory L; Bautista, Cristina; Pong, Erik; Nguyen, Duc-Hanh T; Jacinto, Jonathan; Eivazi, Araz; Muchhal, Umesh S; Karki, Sher; Chu, Seung Y; Lazar, Greg A

2011-01-01

69

Development of a Two-part Strategy to Identify a Therapeutic Human Bispecific Antibody That Inhibits IgE Receptor Signaling*  

PubMed Central

The development of bispecific antibodies as therapeutic agents for human diseases has great clinical potential, but broad application has been hindered by the difficulty of identifying bispecific antibody formats that exhibit favorable pharmacokinetic properties and ease of large-scale manufacturing. Previously, the development of an antibody technology utilizing heavy chain knobs-into-holes mutations and a single common light chain enabled the small-scale generation of human full-length bispecific antibodies. Here we have extended the technology by developing a two-part bispecific antibody discovery strategy that facilitates proof-of-concept studies and clinical candidate antibody generation. Our scheme consists of the efficient small-scale generation of bispecific antibodies lacking a common light chain and the hinge disulfides for proof-of-concept studies coupled with the identification of a common light chain bispecific antibody for large-scale production with high purity and yield. We have applied this technology to generate a bispecific antibody suitable for development as a human therapeutic. This antibody directly inhibits the activation of the high affinity IgE receptor Fc?RI on mast cells and basophils by cross-linking Fc?RI with the inhibitory receptor Fc?RIIb, an approach that has strong therapeutic potential for asthma and other allergic diseases. Our approach for producing human bispecific full-length antibodies enables the clinical application of bispecific antibodies to a validated therapeutic pathway in asthma.

Jackman, Janet; Chen, Yongmei; Huang, Arthur; Moffat, Barbara; Scheer, Justin M.; Leong, Steven R.; Lee, Wyne P.; Zhang, Juan; Sharma, Navneet; Lu, Yanmei; Iyer, Suhasini; Shields, Robert L.; Chiang, Nancy; Bauer, Michele C.; Wadley, Diana; Roose-Girma, Merone; Vandlen, Richard; Yansura, Daniel G.; Wu, Yan; Wu, Lawren C.

2010-01-01

70

A bispecific antibody based assay shows potential for detecting tuberculosis in resource constrained laboratory settings.  

PubMed

The re-emergence of tuberculosis (TB) as a global public health threat highlights the necessity of rapid, simple and inexpensive point-of-care detection of the disease. Early diagnosis of TB is vital not only for preventing the spread of the disease but also for timely initiation of treatment. The later in turn will reduce the possible emergence of multi-drug resistant strains of Mycobacterium tuberculosis. Lipoarabinomannan (LAM) is an important non-protein antigen of the bacterial cell wall, which is found to be present in different body fluids of infected patients including blood, urine and sputum. We have developed a bispecific monoclonal antibody with predetermined specificities towards the LAM antigen and a reporter molecule horseradish peroxidase (HRPO). The developed antibody was subsequently used to design a simple low cost immunoswab based assay to detect LAM antigen. The limit of detection for spiked synthetic LAM was found to be 5.0 ng/ml (bovine urine), 0.5 ng/ml (rabbit serum) and 0.005 ng/ml (saline) and that for bacterial LAM from M. tuberculosis H37Rv was found to be 0.5 ng/ml (rabbit serum). The assay was evaluated with 21 stored clinical serum samples (14 were positive and 7 were negative in terms of anti-LAM titer). In addition, all 14 positive samples were culture positive. The assay showed 100% specificity and 64% sensitivity (95% confidence interval). In addition to good specificity, the end point could be read visually within two hours of sample collection. The reported assay might be used as a rapid tool for detecting TB in resource constrained laboratory settings. PMID:22363820

Sarkar, Susmita; Tang, Xinli L; Das, Dipankar; Spencer, John S; Lowary, Todd L; Suresh, Mavanur R

2012-01-01

71

Redirection of T-cell effector functions for cancer therapy: bispecific antibodies and chimeric antigen receptors.  

PubMed

T cells are the most potent cells of the immune system; however, they fail in the immunosurveillance of tumors. In previous decades, scientists began studying methods to take advantage of T-cell potency in cancer therapy by redirecting them against tumors independently from the T-cell receptor-defined specificity. Among different approaches, the most promising are the use of bispecific antibodies and T-cell engineering to create chimeric antigen receptors. Bispecific antibodies, by simultaneously recognizing target antigen and an activating receptor on the surface of an immune effector cell, offer an opportunity to redirect immune effector cells to kill cancer cells. The other approach is the generation of chimeric antigen receptors by fusing extracellular antibodies to intracellular signaling domains. Chimeric antigen receptor-engineered T cells are able to specifically kill tumor cells in a MHC-independent way. The efficacy of these reagents in different formats has been clinically validated and will be presented here. PMID:23560375

Satta, Alessandro; Mezzanzanica, Delia; Turatti, Fabio; Canevari, Silvana; Figini, Mariangela

2013-04-01

72

Monoclonal antibodies in human cancer.  

PubMed

Mouse, chimeric, humanized and human monoclonal antibodies (MABs) are all in use for treatment of human cancer. Unconjugated antibodies have a complex mechanism of action, dependent on the nature of the target structure. Antibodies can activate the immune system (antibody-dependent cellular cytotoxicity [ADCC], complement-dependent cytotoxicity [CDC], induction of tumor immunity [idiotype network]). ADCC appears to be one of the most important immune effector functions. Antibodies may also induce apoptosis, cell cycle arrest, inhibition of cell proliferation as well as angiogenesis and metastatic spread. For most antibodies there is no clear dose-response relationship in vivo. The effect of antibodies can be enhanced by combination with chemotherapy and/or by agents which activate the immune system. The best therapeutic effect may be obtained if MABs are used early in the course of the disease. Rituximab (anti-CD20) was the first registered MAB for the therapy of follicular lymphoma. Impressive results have been seen in combination with CHOP chemotherapy (cyclophosphamide, doxorubicin, vincristine and prednisone) in follicular and high-grade lymphomas. In other non-Hodgkin's lymphoma subtypes, promising results are also seen in combination with chemotherapy. Trastuzumab (anti-Her2) is a breakthrough in the treatment of breast cancer in combination with chemotherapeutic agents. This antibody is also in clinical testing for adjuvant treatment. Alemtuzumab (anti-CD52) has shown impressive results both in refractory chronic lymphocytic leukemia and as up-front therapy. There are many other antibodies in late stages of testing for registration. Interesting MABs include cetuximab (anti-epidermal growth factor receptor [EGFR]), especially in combination with radiotherapy in head and neck cancer; ABX-EGF (anti-EGFR) in renal carcinoma; bevacizumab (anti-vascular endothelial growth factor) in several solid tumors. Antiepithelial cell adhesion molecule antibodies show promise in combination with chemotherapy in the adjuvant setting of colorectal carcinoma. It is estimated that about 20 antibodies will be in clinical use by the year 2010. PMID:14988743

Mellstedt, H

2003-01-01

73

Monoclonal Antibodies to Cholesterol and Methods.  

National Technical Information Service (NTIS)

This patent application relates to monoclonal antibodies which demonstrate specific reactivity to cholesterol and methods for the detection of high levels of cholesterol by contacting biological specimens containing cholesterol with the monoclonal antibod...

C. R. Alving G. M. Swartz

1986-01-01

74

The role of apoptosis in bispecific antibody-mediated T-cell cytotoxicity.  

PubMed Central

In this report we describe the role of apoptosis in the process of tumour cell killing by bispecific monoclonal antibody (BsMAb)-redirected cytolytic T cells. The BsMAb used, BIS-1, has dual specificity for the CD3 complex on T cells and the pancarcinoma-associated 38 kDa transmembrane antigen EGP-2. BIS-1 allows activated T cells to specifically recognise and kill EGP-2-positive but not EGP-2-negative target cells. An assay was developed to quantify apoptosis in cells by separation of 3H-thymidine-labelled low-molecular, i.e. fragmented, from high-molecular, i.e. non-fragmented DNA. The presence of low molecular weight DNA was measured both within the target cells and in the cell-free supernatant. After exposure to BIS-1-redirected, -activated T cells, apoptosis was observed in EGP-2-positive target cells but not in EGP-2-negative target cells. Also no DNA fragmentation proved to be induced in the activated effector cells during assay. The degree of EGP-2-positive target DNA fragmentation depended on the concentration of BsMAb, the E/T ratio and the incubation time. Using a low E/T ratio (1/1), DNA fragmentation in and 51Cr release from target cells showed similar characteristics and kinetics. At higher E/T ratio (20/1), the 51Cr release from the target cells increased to a greater extent than the percentage fragmented target cell DNA. Inhibitors of DNA fragmentation added to the cytotoxicity assay inhibited not only DNA fragmentation, but also the release of chromium-51 from the target cells, suggesting that apoptosis and cell lysis are closely related in BsMAb-mediated cell killing. Images Figure 1

Kroesen, B. J.; Wellenberg, G. J.; Bakker, A.; Helfrich, W.; The, T. H.; de Leij, L.

1996-01-01

75

Use of trifunctional bispecific antibodies to prevent graft versus host disease induced by allogeneic lymphocytes  

Microsoft Academic Search

A trifunctional bispecific antibody (BiLu) directed against murine CD3 and human epithelial-cell adhesion molecule (Ep- CAM) was tested for its ability to improve cell-mediated adoptive immunotherapy in a murine model of B16 melanoma cells transfected with human EpCAM. Intraperi- toneal inoculation of naive C57BL\\/6 (C57) splenocytes induced lethal graft versus host disease (GVHD) in 85% to 97% of sublethally irradiated

Shoshana Morecki; Horst Lindhofer; Elena Yacovlev; Yael Gelfand; Shimon Slavin

2006-01-01

76

Monoclonal antibody therapeutics with up to five specificities  

PubMed Central

The recognition that few human diseases are thoroughly addressed by mono-specific, monoclonal antibodies (mAbs) continues to drive the development of antibody therapeutics with additional specificities and enhanced activity. Historically, efforts to engineer additional antigen recognition into molecules have relied predominantly on the reformatting of immunoglobulin domains. In this report we describe a series of fully functional mAbs to which additional specificities have been imparted through the recombinant fusion of relatively short polypeptides sequences. The sequences are selected for binding to a particular target from combinatorial libraries that express linear, disulfide-constrained, or domain-based structures. The potential for fusion of peptides to the N- and C- termini of both the heavy and light chains affords the bivalent expression of up to four different peptides. The resulting molecules, called zybodies, can gain up to four additional specificities, while retaining the original functionality and specificity of the scaffold antibody. We explore the use of two clinically significant oncology antibodies, trastuzumab and cetuximab, as zybody scaffolds and demonstrate functional enhancements in each case. The affect of fusion position on both peptide and scaffold function is explored, and penta-specific zybodies are demonstrated to simultaneously engage five targets (ErbB2, EGFR, IGF-1R, Ang2 and integrin ?v?3). Bispecific, trastuzumab-based zybodies targeting ErbB2 and Ang2 are shown to exhibit superior efficacy to trastuzumab in an angiogenesis-dependent xenograft tumor model. A cetuximab-based bispecific zybody that targeting EGFR and ErbB3 simultaneously disrupted multiple intracellular signaling pathways; inhibited tumor cell proliferation; and showed efficacy superior to that of cetuximab in a xenograft tumor model.

LaFleur, David W.; Abramyan, Donara; Kanakaraj, Palanisamy; Smith, Rodger G.; Shah, Rutul R.; Wang, Geping; Yao, Xiao-Tao; Kankanala, Spandana; Boyd, Ernie; Zaritskaya, Liubov; Nam, Viktoriya; Puffer, Bridget A.; Buasen, Pete; Kaithamana, Shashi; Burnette, Andrew F.; Krishnamurthy, Rajesh; Patel, Dimki; Roschke, Viktor V.; Kiener, Peter A.; Hilbert, David M.; Barbas III, Carlos F.

2013-01-01

77

Monoclonal Antibody Therapy for Cancer  

Microsoft Academic Search

\\u000a Since the approval of rituximab (Rituxan®) for the treatment of B-cell non-Hodgkin’s lymphoma (B-NHL) in 1997, nine additional monoclonal antibodies (mAbs) have been\\u000a approved by the FDA for cancer therapy. Currently, more than 1,300 clinical studies registered at ClinicalTrials.gov investigate\\u000a mAb therapy of cancer, including more than 150 phase III clinical trials. In concert with their clinical acceptance, mAbs\\u000a in

Christoph Rader

78

LC-MS characterization and purity assessment of a prototype bispecific antibody.  

PubMed

Bispecific IgG asymmetric (heterodimeric) antibodies offer enhanced therapeutic efficacy, but present unique challenges for drug development. These challenges are related to the proper assembly of heavy and light chains. Impurities such as symmetric (homodimeric) antibodies can arise with improper assembly. A new method to assess heterodimer purity of such bispecific antibody products is needed because traditional separation-based purity assays are unable to separate or quantify homodimer impurities. This paper presents a liquid chromatography-mass spectrometry (LC-MS)-based method for evaluating heterodimeric purity of a prototype asymmetric antibody containing two different heavy chains and two identical light chains. The heterodimer and independently expressed homodimeric standards were characterized by two complementary LC-MS techniques: Intact protein mass measurement of deglycosylated antibody and peptide map analyses. Intact protein mass analysis was used to check molecular integrity and composition. LC-MS(E) peptide mapping of Lys-C digests was used to verify protein sequences and characterize post-translational modifications, including C-terminal truncation species. Guided by the characterization results, a heterodimer purity assay was demonstrated by intact protein mass analysis of pure deglycosylated heterodimer spiked with each deglycosylated homodimeric standard. The assay was capable of detecting low levels (2%) of spiked homodimers in conjunction with co-eluting half antibodies and multiple mass species present in the homodimer standards and providing relative purity differences between samples. Detection of minor homodimer and half-antibody C-terminal truncation species at levels as low as 0.6% demonstrates the sensitivity of the method. This method is suitable for purity assessment of heterodimer samples during process and purification development of bispecific antibodies, e.g., clone selection. PMID:23884083

Woods, R Jeremy; Xie, Michael Hongwei; Von Kreudenstein, Thomas Spreter; Ng, Gordon Y K; Dixit, Surjit B

2013-01-01

79

LC-MS characterization and purity assessment of a prototype bispecific antibody  

PubMed Central

Bispecific IgG asymmetric (heterodimeric) antibodies offer enhanced therapeutic efficacy, but present unique challenges for drug development. These challenges are related to the proper assembly of heavy and light chains. Impurities such as symmetric (homodimeric) antibodies can arise with improper assembly. A new method to assess heterodimer purity of such bispecific antibody products is needed because traditional separation-based purity assays are unable to separate or quantify homodimer impurities. This paper presents a liquid chromatography-mass spectrometry (LC-MS)-based method for evaluating heterodimeric purity of a prototype asymmetric antibody containing two different heavy chains and two identical light chains. The heterodimer and independently expressed homodimeric standards were characterized by two complementary LC-MS techniques: Intact protein mass measurement of deglycosylated antibody and peptide map analyses. Intact protein mass analysis was used to check molecular integrity and composition. LC-MSE peptide mapping of Lys-C digests was used to verify protein sequences and characterize post-translational modifications, including C-terminal truncation species. Guided by the characterization results, a heterodimer purity assay was demonstrated by intact protein mass analysis of pure deglycosylated heterodimer spiked with each deglycosylated homodimeric standard. The assay was capable of detecting low levels (2%) of spiked homodimers in conjunction with co-eluting half antibodies and multiple mass species present in the homodimer standards and providing relative purity differences between samples. Detection of minor homodimer and half-antibody C-terminal truncation species at levels as low as 0.6% demonstrates the sensitivity of the method. This method is suitable for purity assessment of heterodimer samples during process and purification development of bispecific antibodies, e.g., clone selection.

Woods, R. Jeremy; Xie, Michael Hongwei; Von Kreudenstein, Thomas Spreter; Ng, Gordon Y.; Dixit, Surjit B.

2013-01-01

80

Uses of monoclonal antibody 8H9  

DOEpatents

This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a pharmaceutically acceptable carrier. This invention also provides an antibody other than the monoclonal antibody 8H9 comprising the complementary determining regions of monoclonal antibody 8H9 or a derivative thereof, capable of binding to the same antigen as the monoclonal antibody 8H9. This invention provides a substance capable of competitively inhibiting the binding of monoclonal antibody 8H9. This invention also provides an isolated scFv of monoclonal antibody 8H9 or a derivative thereof. This invention also provides the 8H9 antigen. This invention also provides a method of inhibiting the growth of tumor cells comprising contacting said tumor cells with an appropriate amount of monoclonal antibody 8H9 or a derivative thereof.

Cheung, Nai-Kong V

2013-08-06

81

The future of monoclonal antibody technology  

PubMed Central

With the rapid growth of monoclonal antibody-based products, new technologies have emerged for creating modified forms of antibodies, including fragments, conjugates and multi-specific antibodies. We created a database of 450 therapeutic antibodies in development to determine which technologies and indications will constitute the “next generation” of antibody products. We conclude that the antibodies of the future will closely resemble the antibodies that have already been approved for commercial sale.

Zider, Alexander

2010-01-01

82

Monoclonal antibodies for serotyping Leishmania strains.  

PubMed Central

Mouse monoclonal antibodies raised against Leishmania tropica major were found to precipitate with the excreted factor produced by this and other leishmanial species. We suggest that a classification system for Leishmania, based on selective precipitation reactions between monoclonal antibodies and the excreted factor, would remove many ambiguities that currently exist. Images

Greenblatt, C L; Slutzky, G M; de Ibarra, A A; Snary, D

1983-01-01

83

Novel Monoclonal Antibody against Human Platelets.  

National Technical Information Service (NTIS)

The production of a monoclonal antibody against human platelets is described in the invention. Unique features of the described monoclonal antibody are that it belongs to the IgGl subclass; it binds only to activated platelets, recognizing a 148,000 kilod...

H. Gralnick

1989-01-01

84

Improved monoclonal antibodies to halodeoxyuridine  

DOEpatents

The development, method of production, characterization and methods of use of two hybridomas, CIdU-1 (ATCC Accession No. HB-8321) and CIdU-2 (ATCC Accession No. HB-8320), are described. These secrete IgG/sub 1/(K) immunoglobulins that react with halodeoxyuridine (HdU or halodU) such as bromo, chloro, fluoro and iodo deoxyuridine (BrdU, CldU, FdU and IdU), whether these are free in solution or incorporated into single stranded DNA in whole cells. The antibodies do not react with naturally occurring free nucleic acids or with deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) polymers. These antibodies are suitable for use in enzyme immunoassays for free CldU, FdU, IdU and BrdU and for detecting cells with these nucleotides incorporated into them. The monoclonal antibodies are useful in the detection of the sensitivity of tumor cells to specific chemotherapeutic agents, in the measurement of the rate of cellular DNA synthesis, in the measurement of the rate of proliferation of normal and malignant cells and in the detection of HPRT deficiency in cells. 1 tab.

Vanderlaan, M.; Dolbeare, F.A.; Gray, J.W.; Thomas, C.B.

1983-10-18

85

Anti-CD22/CD20 Bispecific antibody with enhanced trogocytosis for treatment of Lupus.  

PubMed

The humanized anti-CD22 antibody, epratuzumab, has demonstrated therapeutic activity in clinical trials of lymphoma, leukemia and autoimmune diseases, treating currently over 1500 cases of non-Hodgkin lymphoma, acute lymphoblastic leukemias, Waldenström's macroglobulinemia, Sjögren's syndrome, and systemic lupus erythematosus. Because epratuzumab reduces on average only 35% of circulating B cells in patients, and has minimal antibody-dependent cellular cytotoxicity and negligible complement-dependent cytotoxicity when evaluated in vitro, its therapeutic activity may not result completely from B-cell depletion. We reported recently that epratuzumab mediates Fc/FcR-dependent membrane transfer from B cells to effector cells via trogocytosis, resulting in a substantial reduction of multiple BCR modulators, including CD22, CD19, CD21, and CD79b, as well as key cell adhesion molecules, including CD44, CD62L, and ?7 integrin, on the surface of B cells in peripheral blood mononuclear cells obtained from normal donors or SLE patients. Rituximab has clinical activity in lupus, but failed to achieve primary endpoints in a Phase III trial. This is the first study of trogocytosis mediated by bispecific antibodies targeting neighboring cell-surface proteins, CD22, CD20, and CD19, as demonstrated by flow cytometry and immunofluorescence microscopy. We show that, compared to epratuzumab, a bispecific hexavalent antibody comprising epratuzumab and veltuzumab (humanized anti-CD20 mAb) exhibits enhanced trogocytosis resulting in major reductions in B-cell surface levels of CD19, CD20, CD21, CD22, CD79b, CD44, CD62L and ?7-integrin, and with considerably less immunocompromising B-cell depletion that would result with anti-CD20 mAbs such as veltuzumab or rituximab, given either alone or in combination with epratuzumab. A CD22/CD19 bispecific hexavalent antibody, which exhibited enhanced trogocytosis of some antigens and minimal B-cell depletion, may also be therapeutically useful. The bispecific antibody is a candidate for improved treatment of lupus and other autoimmune diseases, offering advantages over administration of the two parental antibodies in combination. PMID:24841238

Rossi, Edmund A; Chang, Chien-Hsing; Goldenberg, David M

2014-01-01

86

Anti-CD22/CD20 Bispecific Antibody with Enhanced Trogocytosis for Treatment of Lupus  

PubMed Central

The humanized anti-CD22 antibody, epratuzumab, has demonstrated therapeutic activity in clinical trials of lymphoma, leukemia and autoimmune diseases, treating currently over 1500 cases of non-Hodgkin lymphoma, acute lymphoblastic leukemias, Waldenström’s macroglobulinemia, Sjögren’s syndrome, and systemic lupus erythematosus. Because epratuzumab reduces on average only 35% of circulating B cells in patients, and has minimal antibody-dependent cellular cytotoxicity and negligible complement-dependent cytotoxicity when evaluated in vitro, its therapeutic activity may not result completely from B-cell depletion. We reported recently that epratuzumab mediates Fc/FcR-dependent membrane transfer from B cells to effector cells via trogocytosis, resulting in a substantial reduction of multiple BCR modulators, including CD22, CD19, CD21, and CD79b, as well as key cell adhesion molecules, including CD44, CD62L, and ?7 integrin, on the surface of B cells in peripheral blood mononuclear cells obtained from normal donors or SLE patients. Rituximab has clinical activity in lupus, but failed to achieve primary endpoints in a Phase III trial. This is the first study of trogocytosis mediated by bispecific antibodies targeting neighboring cell-surface proteins, CD22, CD20, and CD19, as demonstrated by flow cytometry and immunofluorescence microscopy. We show that, compared to epratuzumab, a bispecific hexavalent antibody comprising epratuzumab and veltuzumab (humanized anti-CD20 mAb) exhibits enhanced trogocytosis resulting in major reductions in B-cell surface levels of CD19, CD20, CD21, CD22, CD79b, CD44, CD62L and ?7-integrin, and with considerably less immunocompromising B-cell depletion that would result with anti-CD20 mAbs such as veltuzumab or rituximab, given either alone or in combination with epratuzumab. A CD22/CD19 bispecific hexavalent antibody, which exhibited enhanced trogocytosis of some antigens and minimal B-cell depletion, may also be therapeutically useful. The bispecific antibody is a candidate for improved treatment of lupus and other autoimmune diseases, offering advantages over administration of the two parental antibodies in combination.

Rossi, Edmund A.; Chang, Chien-Hsing; Goldenberg, David M.

2014-01-01

87

Monoclonal Antibodies for the Treatment of Cancer  

PubMed Central

Over the past decade, the clinical utility of monoclonal antibodies has been realized and antibodies are now a mainstay for the treatment of cancer. Antibodies have the unique capacity to target and kill tumor cells while simultaneously activating immune effectors to kill tumor cells through the complement cascade or antibody-dependent cellular cytotoxicity (ADCC). This multifaceted mechanism of action combined with target specificity underlies the capacity of antibodies to elicit anti-tumor responses while minimizing the frequency and magnitude of adverse events. This review will focus on mechanisms of action, clinical applications and putative mechanisms of resistance to monoclonal antibody therapy in the context of cancer.

Shuptrine, Casey; Surana, Rishi; Weiner, Louis M.

2012-01-01

88

[Therapeutic efficacy of three bispecific antibodies on rheumatoid arthritis mice models].  

PubMed

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA. PMID:24961102

Li, Qing-Cui; Han, Xiao-Hui; Zhou, Bing; Wang, Wen-Fei; Ren, Gui-Ping; Sun, Cui-Yu; Wu, Qiang; Yu, Yin-Hang; Xu, Li-Ming; Wang, Qiu-Ying; Qi, Jian-Ying; Wei, Yu-Quan; Cao, Hong-Wei; Han, Jun-Yan; Li, De-Shan

2014-03-01

89

Novel bispecific antibodies increase ?? T-cell cytotoxicity against pancreatic cancer cells.  

PubMed

The ability of human ?? T cells from healthy donors to kill pancreatic ductal adenocarcinoma (PDAC) in vitro and in vivo in immunocompromised mice requires the addition of ?? T-cell-stimulating antigens. In this study, we demonstrate that ?? T cells isolated from patients with PDAC tumor infiltrates lyse pancreatic tumor cells after selective stimulation with phosphorylated antigens. We determined the absolute numbers of ?? T-cell subsets in patient whole blood and applied a real-time cell analyzer to measure their cytotoxic effector function over prolonged time periods. Because phosphorylated antigens did not optimally enhance ?? T-cell cytotoxicity, we designed bispecific antibodies that bind CD3 or V?9 on ?? T cells and Her2/neu (ERBB2) expressed by pancreatic tumor cells. Both antibodies enhanced ?? T-cell cytotoxicity with the Her2/V?9 antibody also selectively enhancing release of granzyme B and perforin. Supporting these observations, adoptive transfer of ?? T cells with the Her2/V?9 antibody reduced growth of pancreatic tumors grafted into SCID-Beige immunocompromised mice. Taken together, our results show how bispecific antibodies that selectively recruit ?? T cells to tumor antigens expressed by cancer cells illustrate the tractable use of endogenous ?? T cells for immunotherapy. PMID:24448235

Oberg, Hans-Heinrich; Peipp, Matthias; Kellner, Christian; Sebens, Susanne; Krause, Sarah; Petrick, Domantas; Adam-Klages, Sabine; Röcken, Christoph; Becker, Thomas; Vogel, Ilka; Weisner, Dietrich; Freitag-Wolf, Sandra; Gramatzki, Martin; Kabelitz, Dieter; Wesch, Daniela

2014-03-01

90

Application of 300× enhanced fluorescence on a plasmonic chip modified with a bispecific antibody to a sensitive immunosensor.  

PubMed

The grating substrate covered with a metal layer, a plasmonic chip, and a bispecific antibody can play a key role in the sensitive detection of a marker protein with an immunosensor, because of the provision of an enhanced fluorescence signal and the preparation of a sensor surface densely modified with capture antibody, respectively. In this study, one of the tumor markers, a soluble epidermal growth factor receptor (sEGFR), was selected as the target to be detected. The ZnO- and silver-coated plasmonic chip with precise regularity and the appropriate duty ratio in the periodic structure further enhanced the fluorescence intensity. As for sensor surface modification with capture antibody, a bispecific antibody (anti-sEGFR and anti-ZnO antibody), the concentrated bispecific antibody solution was found to nonlinearly form a surface densely immobilized with antibody, because the binding process of a bispecific antibody to the ZnO surface can be a competitive process with adsorption of phosphate. As a result, the interface on the plasmonic chip provided a 300× enhanced fluorescence signal compared with that on a ZnO-coated glass slide, and therefore sEGFR was found to be quantitatively detected in a wide concentration range from 10 nM to 700 fM on our plasmonic surface. PMID:23945148

Tawa, Keiko; Umetsu, Mitsuo; Nakazawa, Hikaru; Hattori, Takamitsu; Kumagai, Izumi

2013-09-11

91

An EpCAM/CD3 bispecific antibody efficiently eliminates hepatocellular carcinoma cells with limited galectin-1 expression.  

PubMed

There have been several studies suggesting that cancer stem cells (CSCs) contribute to the high rates of recurrence and resistance to therapies observed in hepatocellular carcinoma (HCC). Epithelial cell adhesion molecule (EpCAM) has been demonstrated to be a biomarker of CSCs and a potential therapeutic target in HCC. Here, we prepared two anti-EpCAM monoclonal antibodies (1H8 and 2F2) and an anti-EpCAM bispecific T cell engager (BiTE) 1H8/CD3, which was derived from 1H8, and used them to treat HCC in vitro and in vivo. The results demonstrated that all of the developed anti-EpCAM antibodies specifically bound to EpCAM. Neither anti-EpCAM monoclonal antibody had obvious anti-HCC activities in vitro or in vivo. However, anti-EpCAM BiTE 1H8/CD3 induced strong peripheral blood mononuclear cell-dependent cellular cytotoxicity in Huh-7 and Hep3B cells but not EpCAM-negative SK-Hep-1 cells. Notably, 1H8/CD3 completely inhibited the growth of Huh-7 and Hep3B xenografts in vivo. Treatment of the Huh-7 HCC xenografts with 1H8/CD3 significantly suppressed tumor proliferation and reduced the expression of most CSC biomarkers. Intriguingly, galectin-1 (Gal-1) overexpression inhibited 1H8/CD3-induced lymphocytotoxicity in HCCs while knockdown of Gal-1 increased the lymphocytotoxicity. Collectively, these results indicate that anti-EpCAM BiTE 1H8/CD3 is a promising therapeutic agent for HCC treatment. Gal-1 may contribute to the resistance of HCC cells to 1H8/CD3-induced lysis. PMID:24177984

Zhang, Pengfei; Shi, Bizhi; Gao, Huiping; Jiang, Hua; Kong, Juan; Yan, Jin; Pan, Xiaorong; Li, Kesang; Zhang, Pengwei; Yao, Ming; Yang, Shengli; Gu, Jianren; Wang, Hongyang; Li, Zonghai

2014-02-01

92

Monoclonal antibody-based therapies in the treatment of acute lymphoblastic leukemia.  

PubMed

Recent studies have suggested that pediatric-intensive chemotherapy regimens can improve outcomes in adults with acute lymphoblastic leukemia (ALL) up to the age of 45. Above this age, toxicities increase. Monoclonal antibody-based therapies bring the promise of increased response rates without excessive toxicity. The addition of rituximab to combination chemotherapy has shown encouraging results. Newer monoclonal antibody-based therapies linked to cytotoxic agents show promise. These include inotuzumab ozogamicin, an anti-CD22 antibody linked to calicheamicin that has produced significant single-agent responses in relapsed and refractory ALL. Other monoclonal antibodies linked to plant or bacterial toxins are in earlier stages of development. Blinatumomab is a novel bispecific T-cell engaging antibody that combines single chain antibodies to CD19 and CD3 and brings a T cell in close proximity to a leukemic lymphoblast with resulting redirected lysis. This agent has demonstrated encouraging results in both the minimal residual disease setting and the relapsed/refractory setting. Autologous chimeric antigen receptor cells have shown promising responses in indolent B-cell lymphoid malignancies and are being tested in ALL. Many of these agents have the potential to increase response rates in older adults. Trials of many of these monoclonal antibody-based therapies are in various stages of development in the treatment of newly diagnosed ALL. PMID:23714527

Litzow, Mark R

2013-01-01

93

Single-domain antibody-based and linker-free bispecific antibodies targeting Fc?RIII induce potent antitumor activity without recruiting regulatory T cells.  

PubMed

Antibody-dependent cell-mediated cytotoxicity, one of the most prominent modes of action of antitumor antibodies, suffers from important limitations due to the need for optimal interactions with Fc? receptors. In this work, we report the design of a new bispecific antibody format, compact and linker-free, based on the use of llama single-domain antibodies that are capable of circumventing most of these limitations. This bispecific antibody format was created by fusing single-domain antibodies directed against the carcinoembryonic antigen and the activating Fc?RIIIa receptor to human C? and CH1 immunoglobulin G1 domains, acting as a natural dimerization motif. In vitro and in vivo characterization of these Fab-like bispecific molecules revealed favorable features for further development as a therapeutic molecule. They are easy to produce in Escherichia coli, very stable, and elicit potent lysis of tumor cells by human natural killer cells at picomolar concentrations. Unlike conventional antibodies, they do not engage inhibitory Fc?RIIb receptor, do not compete with serum immunoglobulins G for receptor binding, and their cytotoxic activity is independent of Fc glycosylation and Fc?RIIIa polymorphism. As opposed to anti-CD3 bispecific antitumor antibodies, they do not engage regulatory T cells as these latter cells do not express Fc?RIII. Studies in nonobese diabetic/severe combined immunodeficient gamma mice xenografted with carcinoembryonic antigen-positive tumor cells showed that Fab-like bispecific molecules in the presence of human peripheral blood mononuclear cells significantly slow down tumor growth. This new compact, linker-free bispecific antibody format offers a promising approach for optimizing antibody-based therapies. PMID:23757164

Rozan, Caroline; Cornillon, Amélie; Pétiard, Corinne; Chartier, Martine; Behar, Ghislaine; Boix, Charlotte; Kerfelec, Brigitte; Robert, Bruno; Pèlegrin, André; Chames, Patrick; Teillaud, Jean-Luc; Baty, Daniel

2013-08-01

94

Feeder Cells for Monoclonal Antibody Production.  

National Technical Information Service (NTIS)

Monoclonal antibodies are important tools with a wide variety of diagnostic and therapeutic applications. They are also extremely useful in many biochemical research procedures. The invention relates to a cell line which efficiently supports the growth of...

H. Mischak W. Kolch F. Hofer U. R. Rapp

1990-01-01

95

Monoclonal Antibody That Defines Human Myoepithelium  

NASA Astrophysics Data System (ADS)

We have isolated a mouse monoclonal antibody that, upon immunohistochemical localization in frozen sections, displays specificity for human myoepithelial cells in the resting mammary gland, sweat glands, and salivary glands. Furthermore, this antibody was strongly and homogeneously reactive with frozen sections of 3 of 60 breast carcinoma specimens. Using immunolocalization techniques in conjunction with polyacrylamide gel electrophoresis, we have determined that the reactivity of this monoclonal antibody is directed toward a 51,000-dalton keratin polypeptide. The potential uses of this antibody in the prognosis of human mammary carcinoma and in understanding the role of the myoepithelium in development and differentiation are discussed.

Dairkee, Shahnaz Hashmi; Blayney, Carlene; Smith, Helene S.; Hackett, Adeline J.

1985-11-01

96

A new class of bispecific antibodies to redirect T cells for cancer immunotherapy.  

PubMed

Various constructs of bispecific antibodies (bsAbs) to redirect effector T cells for the targeted killing of tumor cells have shown considerable promise in both preclinical and clinical studies. The single-chain variable fragment (scFv)-based formats, including bispecific T-cell engager (BiTE) and dual-affinity re-targeting (DART), which provide monovalent binding to both CD3 on T cells and to the target antigen on tumor cells, can exhibit rapid blood clearance and neurological toxicity due to their small size (~55 kDa). Herein, we describe the generation, by the modular DOCK-AND-LOCK™) (DNL™) method, of novel T-cell redirecting bispecific antibodies, each comprising a monovalent anti-CD3 scFv covalently conjugated to a stabilized dimer of different anti-tumor Fabs. The potential advantages of this design include bivalent binding to tumor cells, a larger size (~130 kDa) to preclude renal clearance and penetration of the blood-brain barrier, and potent T-cell mediated cytotoxicity. These prototypes were purified to near homogeneity, and representative constructs were shown to provoke the formation of immunological synapses between T cells and their target tumor cells in vitro, resulting in T-cell activation and proliferation, as well as potent T-cell mediated anti-tumor activity. In addition, in vivo studies in NOD/SCID mice bearing Raji Burkitt lymphoma or Capan-1 pancreatic carcinoma indicated statistically significant inhibition of tumor growth compared with untreated controls. PMID:24492297

Rossi, Diane L; Rossi, Edmund A; Cardillo, Thomas M; Goldenberg, David M; Chang, Chien-Hsing

2014-01-01

97

Optimization studies for the coupling of bispecific antibodies to viral anchor molecules of a tumor vaccine.  

PubMed

Tumor vaccines have to provide several signals for T cell activation. Among them, signal 1 (through TCR/CD3) and signal 2 (through CD28) are the most important. We herein describe a procedure to introduce anti-CD3 and anti-CD28 signals into any tumor cell which is susceptible to infection by Newcastle disease virus (NDV). We developed the ATV-NDV tumor vaccine which consists of patient-derived tumor cells (ATV) modified through infection by NDV. We tested for further improvement of vaccine efficiency the addition of two bispecific single-chain antibodies. They bind with one arm to the viral hemagglutinin-neuraminidase (HN) or fusion (F) protein of NDV expressed at the surface of the vaccine cells while the second arm is directed either against CD3 or CD28 of T cells. The aim of this study was to optimize the coupling of these new reagents to the tumor vaccine. When anti-CD3 and anti-CD28 molecules bind to the same anchoring viral molecule (e.g. HN), competition for binding could occur under certain conditions. This was not the case when the bispecific reagents bound to separate viral molecules (HN or F, respectively). When using transfectants expressing HN and F either separately or on the same cell, we show that T cell activation works best when anti-CD3 and anti-CD28 are attached to the same stimulator cell. The clinical application of such a combined therapy with ATV-NDV vaccine cells and bi-specific antibodies allows to modify the strength of signal 1 and 2 in a quantitative and predictable way according to the immune status of the T cells and the requirements of the patients' immune system. PMID:20878068

Fournier, Philippe; Aigner, Maximilian; Schirrmacher, Volker

2010-11-01

98

Optimized monoclonal antibodies against tissue factor pathway inhibitor (TFPI)  

US Patent & Trademark Office Database

Isolated monoclonal antibodies that bind human tissue factor pathway inhibitor (TFPI) are provided. Isolated nucleic acid molecules encoding monoclonal antibodies that bind TFPI are also contemplated. Pharmaceutical compositions comprising the anti-TFPI monoclonal antibodies and methods of treating deficiencies or defects in coagulation by administration of the antibodies are also provided. Methods of producing the antibodies are also provided.

2013-07-09

99

Monoclonal Antibodies to Herpes Simplex Virus Type I Polypeptides.  

National Technical Information Service (NTIS)

It is now possible to make pure antibodies to proteins by application of the 'monoclonal antibody' or 'hybridoma' technique. The present invention is one such new technique which embodies the monoclonal antibody mechanism. About 50 proteins have been disc...

B. Hampar M. Zweig S. D. Showalter

1982-01-01

100

Identification and Multidimensional Optimization of an Asymmetric Bispecific IgG Antibody Mimicking the Function of Factor VIII Cofactor Activity  

PubMed Central

In hemophilia A, routine prophylaxis with exogenous factor VIII (FVIII) requires frequent intravenous injections and can lead to the development of anti-FVIII alloantibodies (FVIII inhibitors). To overcome these drawbacks, we screened asymmetric bispecific IgG antibodies to factor IXa (FIXa) and factor X (FX), mimicking the FVIII cofactor function. Since the therapeutic potential of the lead bispecific antibody was marginal, FVIII-mimetic activity was improved by modifying its binding properties to FIXa and FX, and the pharmacokinetics was improved by engineering the charge properties of the variable region. Difficulties in manufacturing the bispecific antibody were overcome by identifying a common light chain for the anti-FIXa and anti-FX heavy chains through framework/complementarity determining region shuffling, and by pI engineering of the two heavy chains to facilitate ion exchange chromatographic purification of the bispecific antibody from the mixture of byproducts. Engineering to overcome low solubility and deamidation was also performed. The multidimensionally optimized bispecific antibody hBS910 exhibited potent FVIII-mimetic activity in human FVIII-deficient plasma, and had a half-life of 3 weeks and high subcutaneous bioavailability in cynomolgus monkeys. Importantly, the activity of hBS910 was not affected by FVIII inhibitors, while anti-hBS910 antibodies did not inhibit FVIII activity, allowing the use of hBS910 without considering the development or presence of FVIII inhibitors. Furthermore, hBS910 could be purified on a large manufacturing scale and formulated into a subcutaneously injectable liquid formulation for clinical use. These features of hBS910 enable routine prophylaxis by subcutaneous delivery at a long dosing interval without considering the development or presence of FVIII inhibitors. We expect that hBS910 (investigational drug name: ACE910) will provide significant benefit for severe hemophilia A patients.

Sampei, Zenjiro; Igawa, Tomoyuki; Soeda, Tetsuhiro; Okuyama-Nishida, Yukiko; Moriyama, Chifumi; Wakabayashi, Tetsuya; Tanaka, Eriko; Muto, Atsushi; Kojima, Tetsuo; Kitazawa, Takehisa; Yoshihashi, Kazutaka; Harada, Aya; Funaki, Miho; Haraya, Kenta; Tachibana, Tatsuhiko; Suzuki, Sachiyo; Esaki, Keiko; Nabuchi, Yoshiaki; Hattori, Kunihiro

2013-01-01

101

Identification and multidimensional optimization of an asymmetric bispecific IgG antibody mimicking the function of factor VIII cofactor activity.  

PubMed

In hemophilia A, routine prophylaxis with exogenous factor VIII (FVIII) requires frequent intravenous injections and can lead to the development of anti-FVIII alloantibodies (FVIII inhibitors). To overcome these drawbacks, we screened asymmetric bispecific IgG antibodies to factor IXa (FIXa) and factor X (FX), mimicking the FVIII cofactor function. Since the therapeutic potential of the lead bispecific antibody was marginal, FVIII-mimetic activity was improved by modifying its binding properties to FIXa and FX, and the pharmacokinetics was improved by engineering the charge properties of the variable region. Difficulties in manufacturing the bispecific antibody were overcome by identifying a common light chain for the anti-FIXa and anti-FX heavy chains through framework/complementarity determining region shuffling, and by pI engineering of the two heavy chains to facilitate ion exchange chromatographic purification of the bispecific antibody from the mixture of byproducts. Engineering to overcome low solubility and deamidation was also performed. The multidimensionally optimized bispecific antibody hBS910 exhibited potent FVIII-mimetic activity in human FVIII-deficient plasma, and had a half-life of 3 weeks and high subcutaneous bioavailability in cynomolgus monkeys. Importantly, the activity of hBS910 was not affected by FVIII inhibitors, while anti-hBS910 antibodies did not inhibit FVIII activity, allowing the use of hBS910 without considering the development or presence of FVIII inhibitors. Furthermore, hBS910 could be purified on a large manufacturing scale and formulated into a subcutaneously injectable liquid formulation for clinical use. These features of hBS910 enable routine prophylaxis by subcutaneous delivery at a long dosing interval without considering the development or presence of FVIII inhibitors. We expect that hBS910 (investigational drug name: ACE910) will provide significant benefit for severe hemophilia A patients. PMID:23468998

Sampei, Zenjiro; Igawa, Tomoyuki; Soeda, Tetsuhiro; Okuyama-Nishida, Yukiko; Moriyama, Chifumi; Wakabayashi, Tetsuya; Tanaka, Eriko; Muto, Atsushi; Kojima, Tetsuo; Kitazawa, Takehisa; Yoshihashi, Kazutaka; Harada, Aya; Funaki, Miho; Haraya, Kenta; Tachibana, Tatsuhiko; Suzuki, Sachiyo; Esaki, Keiko; Nabuchi, Yoshiaki; Hattori, Kunihiro

2013-01-01

102

An Alternative Chemical Redox Method for the Production of Bispecific Antibodies: Implication in Rapid Detection of Food Borne Pathogens  

PubMed Central

Bi-functional antibodies with the ability to bind two unrelated epitopes have remarkable potential in diagnostic and bio-sensing applications. In the present study, bispecific antibodies that recognize human red blood cell (RBC) and the food borne pathogen Listeria monocytogenes (L. monocytogenes) were engineered. The procedure involves initial reduction of a mixture of anti-RBC and anti-Listeria antibodies followed by gradual re-oxidation of the reduced disulphides. This facilitates association of the separated antibody chains and formation of hybrid immunoglobulins with affinity for the L. monocytogenes and human RBC. The bispecific antibodies caused the agglutination of the RBCs only in the presence of L. monocytogenes cells. The agglutination process necessitated the specific presence of L. monocytogenes and the red colored clumps formed were readily visible with naked eyes. The RBC agglutination assay described here provides a remarkably simple approach for the rapid and highly specific screening of various pathogens in their biological niches.

Owais, Mohammad; Kazmi, Shadab; Tufail, Saba; Zubair, Swaleha

2014-01-01

103

An alternative chemical redox method for the production of bispecific antibodies: implication in rapid detection of food borne pathogens.  

PubMed

Bi-functional antibodies with the ability to bind two unrelated epitopes have remarkable potential in diagnostic and bio-sensing applications. In the present study, bispecific antibodies that recognize human red blood cell (RBC) and the food borne pathogen Listeria monocytogenes (L. monocytogenes) were engineered. The procedure involves initial reduction of a mixture of anti-RBC and anti-Listeria antibodies followed by gradual re-oxidation of the reduced disulphides. This facilitates association of the separated antibody chains and formation of hybrid immunoglobulins with affinity for the L. monocytogenes and human RBC. The bispecific antibodies caused the agglutination of the RBCs only in the presence of L. monocytogenes cells. The agglutination process necessitated the specific presence of L. monocytogenes and the red colored clumps formed were readily visible with naked eyes. The RBC agglutination assay described here provides a remarkably simple approach for the rapid and highly specific screening of various pathogens in their biological niches. PMID:24637674

Owais, Mohammad; Kazmi, Shadab; Tufail, Saba; Zubair, Swaleha

2014-01-01

104

Receptor Monoclonal Antibodies that Inhibit Tumor Angiogenesis.  

National Technical Information Service (NTIS)

The purpose of this project is to generate monoclonal antibodies to receptor molecules for the angiogenic growth factor VEGF. These antibodies will be tested for their potential as inhibitors of VEGF-stimulated angiogenesis, which is thought to play an im...

J. D. Sato

1999-01-01

105

Monoclonal Antibodies in Paediatric Acute Lymphoblastic Leukemia  

Microsoft Academic Search

\\u000a Development of monoclonal antibodies (moAbs) for treatment of haematological malignancies is a rapidly growing field. Whereas\\u000a unconjugated humanized antibodies are well tolerated and may be easily combined with chemotherapy, immunoconjugates delivering\\u000a toxic compounds right into the target cells exhibit more severe side effects. Highly and selectively expressed antigens are\\u000a ideal targets for antibody treatment and suitable for broad development in

Arend von Stackelberg

106

Therapeutic efficacy of three bispecific antibodies on collagen-induced arthritis mouse model.  

PubMed

Interleukin-1? (IL-1?) and interleukin-17A (IL-17A) are inducible factors and important cytokines in the pathogenesis of rheumatoid arthritis (RA). In the present study, three bispecific and neutralizing antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1? and hIL-17A were constructed, their therapeutic efficacy was compared on collagen induced arthritis (CIA) model mice. In vitro assays demonstrated that the three antibodies could simultaneously bind to target both hIL-1? and hIL-17A. Mice with CIA were subcutaneously administered with one of three antibodies every two days for 29days, we noticed that, compared with the BsAB-2 and BsAB-3, BsAB-1 antibody therapy resulted in more significant effect on alleviating the severity of arthritis by preventing bone damage and cartilage destruction and substantially decreasing production of CII-specific antibodies. In addition, BsAB-1 antibody was more potent in the inhibition of mRNA expression of IL-2, IL-1?, IL-17A, TNF-? and MMP-3 in the spleen of CIA mice compared to the other two. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 for the treatment of RA model mice, and may be chosen as an ideal candidate for further development of therapeutic drugs for treatment of RA. PMID:24800661

Li, Qingcui; Ren, Guiping; Xu, Liming; Wang, Qiuying; Qi, Jianying; Wang, Wenfei; Zhou, Bing; Han, Xiaohui; Sun, Cuiyu; Wu, Qiang; Yu, Yinhang; Peng, Zhongyi; Zheng, Shimin; Li, Deshan

2014-07-01

107

Radioimmunodetection of medullary thyroid cancer using a bispecific anti-CEA/anti-indium-DTPA antibody and an indium-111-labeled DTPA dimer.  

PubMed

Two-step radioimmunotargeting using a bispecific anti-CEA/anti-in-DTPA monoclonal antibody and an 111In-labeled DTPA dimer (diDTPA-TL) was evaluated nine times in eight patients with medullary thyroid cancer (MTC). Immunoscintigraphy was performed 5 and 24 hr after injection of 111In-diDTPA-TL. For five patients, radioimmunoguided surgery (RIGS) was performed using a hand-held gamma probe (sodium iodine), and a biodistribution study was performed 48 hr (four times) and 24 hr (one time) after injection of 111In-diDTPA-TL. Mean tumor uptake (%ID/kg in tumor) was 39 (range 2.75-139). In these five patients, immunoscintigraphy visualized all known tumors and detected unknown foci (US and CT were negative) in the neck (once) and neck and liver (once). Immunoscintigraphy, performed four times in search of a recurrence, detected unknown localizations in the mediastinum and neck (twice) and was negative twice. There were no false-positives. In three of five patients who had surgery, RIGS localized tumor foci not detected by the surgeon. RIGS failed to detect two small lesions (10 x 10 mm) corresponding to sites of fibrosis and microscopic cancer infiltration. Bispecific anti-CEA/anti-In-DTPA mediated targeting of 111In-diDTPA-TL provided elevated tumor uptake and tumor-to-normal tissue ratios. Radioimmunodetection of small MTC lesions is thus possible even when morphological imaging techniques prove negative. PMID:8326383

Peltier, P; Curtet, C; Chatal, J F; Le Doussal, J M; Daniel, G; Aillet, G; Gruaz-Guyon, A; Barbet, J; Delaage, M

1993-08-01

108

Production of human monoclonal antibodies to myeloperoxidase.  

PubMed Central

Two mouse-human heterohybridomas secreting human antibodies to myeloperoxidase (MPO) were derived from the peripheral blood of a patient who developed microscopic polyarteritis as the result of long-term treatment with hydralazine. Forty-five immunoglobulin-secreting lines were obtained from the fusion of patient lymphocytes with the CB-F7 heteromyeloma cell line. Of these, two antibodies, one IgG and one IgM, bound to myeloperoxidase in solid phase ELISA and gave a perinuclear staining pattern on ethanol-fixed human neutrophil cytospin preparations. The staining patterns were similar to those seen with serum from the patient. Antigen-inhibition studies revealed that the affinity of the IgG monoclonal antibody was 28 times higher (k = 1.4 x 10(-7)) than the IgM antibody (k = 5 x 10(-5)). Cross-inhibition studies further suggested that the two monoclonal antibodies recognized the same epitope on MPO. Of the other secreting cell lines, none produced antibody which reacted with the panel of autoantigens used for testing. Neither mononuclear antibody reacted with this panel indicating that they were not simply polyreactive natural autoantibodies. These are the first human monoclonal antibodies to native myeloperoxidase to be reported. Images Figure 2 Figure 3

Ehrenstein, M R; Leaker, B; Isenberg, D; Cambridge, G

1992-01-01

109

Rabbit monoclonal antibody: potential application in cancer therapy  

PubMed Central

By targeting antigens specifically, monoclonal antibodies represent a new class of therapeutic agents for the clinical management of various diseases including cancers. Monoclonal antibody technology has been greatly developed by reducing murine content in antibodies to minimize side effects in clinical applications. However, several intrinsic disadvantages of antibodies with murine origin limit the clinical efficacy of monoclonal antibodies based targeted therapy. The development of rabbit monoclonal antibody technology provides an alternative source of monoclonal antibodies with higher specificity and less cost for the development of routine targeted therapy against cancers.

Feng, Lifeng; Wang, Xian; Jin, Hongchuan

2011-01-01

110

[Use of monoclonal antibodies in HLA immunology].  

PubMed

The author gives an account of the classification and use of HLA monoclonal antibodies. In experimental work the author demonstrates the contribution of investigations of the expressivity of histocompatible antigens class I and II and their changes in sound and tumourous tissues. In clinical practice they are important in particular in transplantology--cleaning of bone marrow, follow up of the GvH reaction, monitoring of patients. HLA monoclonal antibodies serve to reveal the polymorphism in the HLA sphere and the follow up of general biological laws. PMID:2327089

Prazák, J

1990-01-01

111

A novel bispecific single-chain antibody for ADAM17 and CD3 induces T-cell-mediated lysis of prostate cancer cells.  

PubMed

ADAM17 (A disintegrin and metalloproteinase 17) is a membrane-bound protease that cleaves various cell surface proteins, including cytokines and cytokine receptors. Recently it was shown that ADAM17 is highly expressed on the surface of many cancer cells, whereas normal cells express low levels of ADAM17, implying that ADAM17 is a potential immunotherapeutic target. We have generated a monoclonal antibody against human ADAM17, which recognized the membrane proximal cysteine-rich extension of the ADAM17 protein. Unlike normal cells, tumour cell lines, such as a prostate cancer cell line, pancreatic cancer cell lines, a breast cancer cell line and a non-small lung cancer cell line, expressed ADAM17 on the cell surface. Using the sequence of the antibody we generated an ADAM17-specific scFv (single-chain variable fragment) and fused this to a CD3-specific scFv to generate a bispecific T-cell engager antibody [A300E-BiTE (bispecific T-cell engager antibody)]. Specificity was demonstrated on cells in which ADAM17 was knocked down with a specific shRNA (short hairpin RNA). A300E-BiTE recognized ADAM17 and CD3 on the cell surface of tumour cells and T-cells respectively. In the presence of primary human peripheral blood mononuclear cells or human T-cells the addition of A300E-BiTE led to ADAM17-specific killing of prostate tumour cells indicating a novel strategy for the treatment of cancer. PMID:22509934

Yamamoto, Kosuke; Trad, Ahmad; Baumgart, Anja; Hüske, Linda; Lorenzen, Inken; Chalaris, Athena; Grötzinger, Joachim; Dechow, Tobias; Scheller, Jürgen; Rose-John, Stefan

2012-07-01

112

Monoclonal antibodies for the treatment of asthma.  

PubMed

Asthma is a chronic inflammatory disease of the airways which can have a detrimental effect on quality of life and in extreme cases cause death. Although the majority of patients can control their asthma symptoms with a combination of steroids and beta agonists there is still a group of patients whose asthma remains symptomatic despite the best available treatment. These severe asthmatic patients represent the unmet medical need in asthma and are the focus of those developing novel monoclonal antibody based drugs. The complex networks of cytokines and cells involved in the pathology of asthma provide plenty of scope for intervention with monoclonal antibody based drugs which are able to block cytokine or chemokine receptor interactions, deplete cells expressing a specific receptor or block cell/cell interactions. At present anti-IgE (Xolair©) is the only monoclonal antibody based drug approved for the treatment of asthma. However, a number of other antibody based drugs have been clinically tested in asthma including anti-IL-5, anti-IL-4, anti-IL-13, anti-TNF?, anti-CCR3, anti-CCR4 and anti-OX40L. This review will examine the development of these monoclonal antibody based therapies. Since many of these therapies have targeted key pathways in asthma pathology these studies provide information on patient stratification and asthma pathology. PMID:21944943

Catley, Matthew C; Coote, Julie; Bari, Mohamed; Tomlinson, Kate L

2011-12-01

113

5th Annual Monoclonal Antibodies Conference  

PubMed Central

The conference, which was organized by Visiongain and held at the BSG Conference Center in London, provided an excellent opportunity for participants to exchange views on the development, production and marketing of therapeutic antibodies, and discuss the current business environment. The conference included numerous interactive panel and group discussions on topics such as isotyping for therapeutic antibodies (panel chair: Nick Pullen, Pfizer), prospects for fully human monoclonal antibodies (chair: Christian Rohlff, Oxford BioTherapeutics), perspectives on antibody manufacturing and development (chair: Bo Kara, Avecia), market impact and post-marketing issues (chair: Keith Rodgers, Bodiam Consulting) and angiogenesis inhibitors (chair: David Blakey, AstraZeneca).

2009-01-01

114

Preparation of Human Monoclonal Antibodies of Selected Specificity and Isotypes.  

National Technical Information Service (NTIS)

Human monoclonal antibodies of predetermined specificity are produced by isolating human B lymphocytes of a given specificity and transforming them with Epstein-Barr Virus. Similarly, human monoclonal antibodies of a predetermined class can be produced by...

P. Casali A. L. Notkins

1987-01-01

115

Characterization and utilization of a monoclonal antibody against pancreatic carcinoma.  

National Technical Information Service (NTIS)

A monoclonal antibody was produced against a human pancreatic adenocarcinoma line and was found to react with several different human carcinomas by immunoperoxidase staining of fixed tissues. The original cells used to generate the monoclonal antibody wer...

S. H. Kurtzman W. F. Sindelar R. W. Atcher J. B. Mitchell W. G. DeGraff

1994-01-01

116

Systemic administration of a bispecific antibody targeting EGFRvIII successfully treats intracerebral glioma.  

PubMed

Bispecific antibodies (bscAbs), particularly those of the bispecific T-cell engager (BiTE) subclass, have been shown to effectively redirect T cells against cancer. Previous efforts to target antigens expressed in both tumors and normal tissues have produced significant toxicity, however. Moreover, like other large molecules, bscAbs may be restricted from entry into the "immunologically privileged" CNS. A tumor-specific mutation of the epidermal growth factor receptor, EGFRvIII, is a constitutively activated tyrosine kinase not found in normal tissues but frequently expressed in glioblastomas and many other neoplasms. Because it is localized solely to tumor tissue, EGFRvIII presents an ideal target for immunotherapy. Here we report the preclinical evaluation of an EGFRvIII-targeted BiTE, bscEGFRvIIIxCD3. Our results show that bscEGFRvIIIxCD3 activates T cells to mediate potent and antigen-specific lysis of EGFRvIII-expressing gliomas in vitro (P < 0.001) at exceedingly low concentrations (10 ng/mL) and effector-to-target ratios (2.5:1). Treatment with i.v. bscEGFRvIIIxCD3 yielded extended survival in mice with well-established intracerebral tumors (P < 0.05) and achieved durable complete cure at rates up to 75%. Antitumor efficacy was significantly abrogated on blockade of EGFRvIII binding, demonstrating the need for target antigen specificity both in vitro and in vivo. These results demonstrate that BiTEs can be used to elicit functional antitumor immunity in the CNS, and that peptide blockade of BiTE-mediated activity may greatly enhance the safety profile for antibody-redirected T-cell therapies. Finally, bscEGFRvIIIxCD3 represents a unique advancement in BiTE technology given its exquisite tumor specificity, which enables precise elimination of cancer without the risk of autoimmune toxicity. PMID:23248284

Choi, Bryan D; Kuan, Chien-Tsun; Cai, Mingqing; Archer, Gary E; Mitchell, Duane A; Gedeon, Patrick C; Sanchez-Perez, Luis; Pastan, Ira; Bigner, Darell D; Sampson, John H

2013-01-01

117

Systemic administration of a bispecific antibody targeting EGFRvIII successfully treats intracerebral glioma  

PubMed Central

Bispecific antibodies (bscAbs), particularly those of the bispecific T-cell engager (BiTE) subclass, have been shown to effectively redirect T cells against cancer. Previous efforts to target antigens expressed in both tumors and normal tissues have produced significant toxicity, however. Moreover, like other large molecules, bscAbs may be restricted from entry into the “immunologically privileged” CNS. A tumor-specific mutation of the epidermal growth factor receptor, EGFRvIII, is a constitutively activated tyrosine kinase not found in normal tissues but frequently expressed in glioblastomas and many other neoplasms. Because it is localized solely to tumor tissue, EGFRvIII presents an ideal target for immunotherapy. Here we report the preclinical evaluation of an EGFRvIII-targeted BiTE, bscEGFRvIIIxCD3. Our results show that bscEGFRvIIIxCD3 activates T cells to mediate potent and antigen-specific lysis of EGFRvIII-expressing gliomas in vitro (P < 0.001) at exceedingly low concentrations (10 ng/mL) and effector-to-target ratios (2.5:1). Treatment with i.v. bscEGFRvIIIxCD3 yielded extended survival in mice with well-established intracerebral tumors (P < 0.05) and achieved durable complete cure at rates up to 75%. Antitumor efficacy was significantly abrogated on blockade of EGFRvIII binding, demonstrating the need for target antigen specificity both in vitro and in vivo. These results demonstrate that BiTEs can be used to elicit functional antitumor immunity in the CNS, and that peptide blockade of BiTE-mediated activity may greatly enhance the safety profile for antibody-redirected T-cell therapies. Finally, bscEGFRvIIIxCD3 represents a unique advancement in BiTE technology given its exquisite tumor specificity, which enables precise elimination of cancer without the risk of autoimmune toxicity.

Choi, Bryan D.; Kuan, Chien-Tsun; Cai, Mingqing; Archer, Gary E.; Mitchell, Duane A.; Gedeon, Patrick C.; Sanchez-Perez, Luis; Pastan, Ira; Bigner, Darell D.; Sampson, John H.

2013-01-01

118

Monoclonal antibodies to human malignant mesothelioma  

Microsoft Academic Search

Murine monoclonal antibodies were used to identify tumor-cell membrane antigens on a new human mesothelioma cell line. Hybridomas were constructed by fusing SP2\\/0 mouse myeloma cells with spleen cells from Balb\\/C mice immunized by the human mesothelioma cell line MT-1. Hybridoma antibody was detected in 55\\/672 microculture wells that reacted to these MT-1 tumor cells by an indirect125I-protein A binding

Timothy M. Anderson; E. Carmack Holmes; Calvin J. Kosaka; Lorna Cheng; Romaine E. Saxton

1987-01-01

119

A Monoclonal Antibody to Limbic System Neurons  

Microsoft Academic Search

A monoclonal antibody produced against hippocampal cell membranes labeled the surface of neurons in the rat limbic system. With a few exceptions, all nonlimbic components were unstained. This specific distribution of immunopositive neurons provides strong evidence of molecular specificity among functionally related neurons in the mammalian brain and supports the concept of a limbic system.

Pat Levitt

1984-01-01

120

Subcutaneous Administration of Monoclonal Antibodies in Oncology  

PubMed Central

Treatment with monoclonal antibodies (mabs) has become an established component of oncological therapy. The monoclonal antibodies available for this purpose are mainly administered intravenously in individually adapted doses according to body weight over longer treatment times. For other chronic diseases such as, for example, diabetes mellitus, the subcutaneous administration of drugs is an established therapy option. For the subcutaneous administration of larger volumes as needed for mab solutions the extracellular matrix of the subcutaneous tissue represents a problem. The co-formulation with recombinant human hyaluronidase makes the relatively pain-free administration of larger fluid volumes and thus the subcutaneous administration of monoclonal antibodies possible, as illustrated by the development of a subcutaneous formulation of trastuzumab. This constitutes a less invasive, time-optimised and flexible form of administration for patients with HER2-positive breast cancer that, with its fixed dosing possibilities, contributes to therapeutic safety. The example of trastuzumab shows that the subcutaneous administration of monoclonal antibodies can simplify oncological long-term therapy not only for the patients but also for the medical personnel.

Jackisch, C.; Muller, V.; Maintz, C.; Hell, S.; Ataseven, B.

2014-01-01

121

Production of Human Monoclonal Antibodies by Heterohybridomas.  

National Technical Information Service (NTIS)

The goal is to establish an industrial process for producing human monoclonal antibodies (h-MoAb). The authors have already established a hybridoma system between human-mouse hybridomas and human lymphocytes (m.h-h) for producing h-MoAb (presented at the ...

K. Kitano Y. Shintani S. Sasai K. Tsukamoto

1985-01-01

122

Immunostimulatory monoclonal antibodies for cancer therapy  

Microsoft Academic Search

Increasing immune responses with immunostimulatory monoclonal antibodies (mAbs) directed to immune-receptor molecules is a new and exciting strategy in cancer therapy. This expanding class of agents functions on crucial receptors, either antagonizing those that suppress immune responses or activating others that amplify immune responses. Complications such as autoimmunity and systemic inflammation are problematic side effects associated with these agents. However,

Sandra Hervas-Stubbs; Martin Glennie; Drew M. Pardoll; Ignacio Melero; Lieping Chen

2007-01-01

123

Perspectives on Anti-HER Monoclonal Antibodies  

Microsoft Academic Search

The ability of Herceptin® to prolong survival in women with HER2-overexpressing breast tumors has proven the concept of using humanized or chimeric monoclonal antibodies (MAbs) for cancer therapy. MAbs have been developed that are specific for many tumorigenic molecules and receptors. They can potentially be used to treat a range of solid tumors. Among the most promising targets for therapy

Malcolm Ranson; Mark X. Sliwkowski

2002-01-01

124

Database on monoclonal antibodies to cytokeratins.  

PubMed

Cytokeratins (CK) are being extensively used as diagnostic markers for various malignancies and other diseases, including human oral precancer and cancer, due to their tissue specific expression. CK are epithelia specific intermediate filament (IF) proteins, which are expressed in a differentiation dependent and tissue specific manner. There are about 30 polypeptides of CK expressed by different human epithelia. Each type of epithelium expresses about 4-6 polypeptides. CK polypeptides share many common epitopes, due to which the antibodies developed against CK tend to cross react. Therefore, a large number of monoclonal and polyclonal antibodies have been developed to distinguish among these proteins. Many of these antibodies are not only monospecific but are also epitope specific. These antibodies are being used in pathology laboratories for routine diagnosis using immunohistochemistry. A number of fixatives are used for fixation of tissue sections prior to the use of these antibodies. Sometimes, this leads in epitope masking. Hence, it becomes necessary to use a battery of monoclonal antibodies (MAb) for accurate diagnosis. Apart from the use of these antibodies in diagnostics, they are also being used in basic research for the study of CK function and their interactions with associated proteins and membrane proteins. In the present communication an effort has been made to make a comprehensive list of MAb to CK giving information like cross-reactivity, epitope specificity, various fixatives used, etc. along with the source of the antibodies, which will provide useful information to the users. PMID:14747055

Upasani, Ojaswini S; Vaidya, Milind M; Bhisey, Avinash N

2004-03-01

125

Phase Separation in Solutions of Monoclonal Antibodies  

NASA Astrophysics Data System (ADS)

We report the observation of liquid-liquid phase separation (LLPS) in a solution of humanized monoclonal antibodies, IgG2, and the effects of human serum albumin, a major blood protein, on this phase separation. We find a significant reduction of phase separation temperature in the presence of albumin, and a preferential partitioning of the albumin into the antibody-rich phase. We provide a general thermodynamic analysis of the antibody-albumin mixture phase diagram and relate its features to the magnitude of the effective inter-protein interactions. Our analysis suggests that additives (HSA in this report), which have moderate attraction with antibody molecules, may be used to forestall undesirable protein condensation in antibody solutions. Our findings are relevant to understanding the stability of pharmaceutical solutions of antibodies and the mechanisms of cryoglobulinemia.

Benedek, George; Wang, Ying; Lomakin, Aleksey; Latypov, Ramil

2012-02-01

126

Human Monoclonal Antibody Against Mesothelin  

Cancer.gov

Mesothelin is a cell surface protein that is naturally expressed at very low levels, but that is significantly increased in aggressive tumors such as mesotheliomas, and pancreatic and ovarian tumors. Therefore, mesothelin is an excellent candidate for tumor-targeted immunotherapeutics. However, the only antibodies against mesothelin that are currently available for clinical trials are of murine origin. The use of these antibodies may be limited by their potential to elicit adverse immune responses in patients with repeated doses.

127

Monoclonal antibodies against the aster yellows agent.  

PubMed

Hybridoma clones secreting specific monoclonal antibodies against the aster yellows agent, a mycoplasma-like organism, were produced by using partially purified salivary gland preparations from infected leafhopper vectors as the immunogen. After 3947 hybridomas from 20 independent fusions were screened for specific antibody against the aster yellows agent, two table clones were obtained. With these monoclonal antibodies the aster yellows agent in diseased lettuce, periwinkles, and inoculative insects was specifically identified by enzyme-linked immunosorbent assay. The aster yellows agent was serologically differentiated from the mycoplasma-like organisms associated with ash yellows, loofah witches'-broom, paulownia witches'-broom, sweet potato witches'-broom, peanut rosette, maize bushy stunt, and elm phloem necrosis. PMID:17757867

Lin, C P; An Chen, T

1985-03-01

128

Development of humanized bispecific antibodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protooncogene  

Microsoft Academic Search

Summary The HER2 protooncogene encodes a 185-kD transmembrane phosphoglycoprotein, human epidermal growth factor receptor 2 (p185u~Rz), whose amplified expression on the cell surface can lead to malignant transformation. Overexpression of HER2\\/p185 urR2 is strongly correlated with progression of human ovarian and breast carcinomas. Recent studies have shown that human T cells can be targeted with bispecific antibody to react against

M. Refaat Shalaby; H. Michael Shepard; Len Presta; Maria L. Rodrigues; S Peter; C. L. Beverley; Marc Feldmann; Paul Carters

1992-01-01

129

Induction of regular cytolytic T cell synapses by bispecific single-chain antibody constructs on MHC class I-negative tumor cells  

Microsoft Academic Search

Certain bispecific single-chain antibody constructs exhibit an extraordinary potency for polyclonal T cell engagement and target cell lysis. Here we studied the structural basis for this potency, using laser scanning confocal microscopy. Cytolytic human T cell synapses could be triggered either by addition of a specific peptide antigen or an Ep-CAM-\\/CD3-bispecific T cell engager (BiTE). Both kinds of synapses showed

Sonja Offner; Robert Hofmeister; Andrea Romaniuk; Peter Kufer; Patrick A. Baeuerle

2006-01-01

130

Cytosine arabinoside promotes cytotoxic effect of T cells on leukemia cells mediated by bispecific antibody.  

PubMed

Chemotherapeutic drugs can enhance an immune response of the host against the tumor in addition to killing cancer cells by direct cytotoxicity. Therefore, the combination of chemotherapy and immunotherapy is a promising approach for eliminating tumors, particularly in advanced stages. A strategic medication is to use a bispecific antibody format that is capable of recruiting polyclonal T cells around antibody-target-expressing tumor cells. Recently, we have constructed a bispecific antibody, anti-CD3×anti-CD19, in a diabody configuration. In this study, we measured B7 family members B7.1 (CD80) and B7.2 (CD86) expressed on a CD19(+) human leukemia cell line, Nalm-6, stimulated by cytosine arabinoside (Ara-C). We found that a low concentration of Ara-C could upregulate CD80 expressed on CD19(+) Nalm-6 cells. The cytotoxicity of T lymphocytes against Nalm-6 cells in vitro and in vivo mediated by the anti-CD3×anti-CD19 diabody with or without a low dose of Ara-C was compared. The combination of the anti-CD3×anti-CD19 diabody and Ara-C showed the greatest effectiveness in enhancing the cytotoxicity of T cells against the tumor cells in vitro and in vivo. Activated T cells expressed higher levels of CD25 and CD69 and released more interleukin 2. Both perforin/granzyme B system and Fas/FasL pathway were involved in the diabody-induced T-cell cytotoxicity. Moreover, the activated T cells could upregulate ICAM-3 expression on Nalm-6 cells, and inhibition of LFA-1-ICAM-3 interaction impaired cytotoxicity of T cells. It was noted that Ara-C could upregulate CD80 expressed on two of five specimens of acute B lymphoblastic leukemia patient-derived cells. Cytotoxicity of T cells against these two patient-derived cells was enhanced in the presence of the anti-CD3×anti-CD19 diabody. These findings indicate that treatment strategy using both cytotoxic lymphocyte-based immunotherapy and chemotherapy may have synergistic effects. PMID:23879717

Li, Wei; Fan, DongMei; Yang, Ming; Yan, Yan; Shi, RuiZan; Cheng, JunPing; Li, ZhenZhen; Zhang, MengNan; Wang, JianXiang; Xiong, Dongsheng

2013-08-01

131

A novel bispecific EGFR/Met antibody blocks tumor-promoting phenotypic effects induced by resistance to EGFR inhibition and has potent antitumor activity  

PubMed Central

Simultaneous targeting of epidermal growth factor receptor (EGFR) and Met in cancer therapy is under pre-clinical and clinical evaluation. Here, we report the finding that treatment with EGFR inhibitors of various tumor cells, when stimulated with hepatocyte growth factor (HGF) and EGF, results in transient upregulation of phosphorylated AKT. Furthermore, EGFR inhibition in this setting stimulates a pro-invasive phenotype as assessed in Matrigel-based assays. Simultaneous treatment with AKT and EGFR inhibitors abrogates this invasive growth, hence functionally linking signaling and phenotype. This observation implies that during treatment of tumors a balanced ratio of EGFR and Met inhibition is required. To address this, we designed a bispecific antibody targeting EGFR and Met, which has the advantage of a fixed 2:1 stoichiometry. This bispecific antibody inhibits proliferation in tumor cell cultures and co-cultures with fibroblasts in an additive manner compared with treatment with both single agents. In addition, cell migration assays reveal a higher potency of the bispecific antibody in comparison with the antibodies' combination at low doses. We demonstrate that the bispecific antibody inhibits invasive growth, which is specifically observed with cetuximab. Finally, the bispecific antibody potently inhibits tumor growth in a non-small cell lung cancer xenograft model bearing a strong autocrine HGF-loop. Together, our findings strongly support a combination treatment of EGFR and Met inhibitors and further evaluation of resistance mechanisms to EGFR inhibition in the context of active Met signaling.

Castoldi, R; Ecker, V; Wiehle, L; Majety, M; Busl-Schuller, R; Asmussen, M; Nopora, A; Jucknischke, U; Osl, F; Kobold, S; Scheuer, W; Venturi, M; Klein, C; Niederfellner, G; Sustmann, C

2013-01-01

132

A novel bispecific EGFR/Met antibody blocks tumor-promoting phenotypic effects induced by resistance to EGFR inhibition and has potent antitumor activity.  

PubMed

Simultaneous targeting of epidermal growth factor receptor (EGFR) and Met in cancer therapy is under pre-clinical and clinical evaluation. Here, we report the finding that treatment with EGFR inhibitors of various tumor cells, when stimulated with hepatocyte growth factor (HGF) and EGF, results in transient upregulation of phosphorylated AKT. Furthermore, EGFR inhibition in this setting stimulates a pro-invasive phenotype as assessed in Matrigel-based assays. Simultaneous treatment with AKT and EGFR inhibitors abrogates this invasive growth, hence functionally linking signaling and phenotype. This observation implies that during treatment of tumors a balanced ratio of EGFR and Met inhibition is required. To address this, we designed a bispecific antibody targeting EGFR and Met, which has the advantage of a fixed 2:1 stoichiometry. This bispecific antibody inhibits proliferation in tumor cell cultures and co-cultures with fibroblasts in an additive manner compared with treatment with both single agents. In addition, cell migration assays reveal a higher potency of the bispecific antibody in comparison with the antibodies' combination at low doses. We demonstrate that the bispecific antibody inhibits invasive growth, which is specifically observed with cetuximab. Finally, the bispecific antibody potently inhibits tumor growth in a non-small cell lung cancer xenograft model bearing a strong autocrine HGF-loop. Together, our findings strongly support a combination treatment of EGFR and Met inhibitors and further evaluation of resistance mechanisms to EGFR inhibition in the context of active Met signaling. PMID:23812422

Castoldi, R; Ecker, V; Wiehle, L; Majety, M; Busl-Schuller, R; Asmussen, M; Nopora, A; Jucknischke, U; Osl, F; Kobold, S; Scheuer, W; Venturi, M; Klein, C; Niederfellner, G; Sustmann, C

2013-12-12

133

Recent developments in monoclonal antibody radiolabeling techniques  

SciTech Connect

Monoclonal antibodies (MAbs) have shown the potential to serve as selective carriers of radionuclides to specific in vivo antigens. Accordingly, there has been an intense surge of research activity in an effort to develop and evaluate MAb-based radiopharmaceuticals for tumor imaging (radioimmunoscintigraphy) and therapy (radioimmunotherapy), as well as for diagnosing nonmalignant diseases. A number of problems have recently been identified, related to the MAbs themselves and to radiolabeling techniques, that comprise both the selectivity and the specificity of the in vivo distribution of radiolabeled MAbs. This paper will address some of these issues and primarily discuss recent developments in the techniques for radiolabeling monoclonal antibodies that may help resolve problems related to the poor in vivo stability of the radiolabel and may thus produce improved biodistribution. Even though many issues are identical with therapeutic radionuclides, the discussion will focus mainly on radioimmunoscintigraphic labels. 78 refs., 6 tabs.

Srivastava, S.C.; Mease, R.C.

1989-01-01

134

Calculation of Monoclonal Antibody Affinity Constants Directly from Antibody Dilution Curves.  

National Technical Information Service (NTIS)

A method for estimating the affinity constants of monoclonal antibody directly from antibody dilution curves is described. Antibody affinity is a major determinant of the interaction of antigen with antibody. Knowledge of antibody affinity, as well as spe...

W. R. Griswold D. P. Nelson

1985-01-01

135

Monoclonal antibodies to carcino-embryonic antigen  

Microsoft Academic Search

Several approaches were made to produce new monoclonal antibodies (MoAb) to human colonic tumours using different immunogens, such as colon cell lines, and tissue and serum from patients with colon cancer. Six MoAb were produced but all were found lo be reactive with carcino-embryonic antigen (CEA). Through tissue reactivity and competitive inhibition studies it was found that four epitopes could

Jin-Ghee Teh; Ian FC McKenzie; J. G. Teh

1990-01-01

136

Monoclonal antibody targets, kills leukemia cells  

Cancer.gov

Researchers at the University of California, San Diego Moores Cancer Center have identified a humanized monoclonal antibody that targets and directly kills chronic lymphocytic leukemia (CLL) cells. The findings, published in the online Early Edition of the Proceedings of the National Academy of Sciences on March 25, 2013 represent a potential new therapy for treating at least some patients with CLL, the most common type of blood cancer in the United States.

137

Monoclonal Antibody Strategies for Targeting HER2  

Microsoft Academic Search

The HER family is composed of four receptors, HER1 to HER4, is dysregulated, and\\/or shows abnormal signaling activity in a\\u000a broad range of human tumors. The essential role of HER2 in the HER signaling network led to the development of anti-HER2 monoclonal\\u000a antibodies (mAb) for cancer therapy. In particular, the humanized antibody trastuzumab (Herceptin™) has antitumor activity\\u000a against HER2-overexpressing breast

Joan Albanell; Jeffrey S. Ross; Linda Pronk; Pere Gascon

138

Trial watch: Monoclonal antibodies in cancer therapy.  

PubMed

During the past 20 years, dozens-if not hundreds-of monoclonal antibodies have been developed and characterized for their capacity to mediate antineoplastic effects, either as they activate/enhance tumor-specific immune responses, either as they interrupt cancer cell-intrinsic signal transduction cascades, either as they specifically delivery toxins to malignant cells or as they block the tumor-stroma interaction. Such an intense research effort has lead to the approval by FDA of no less than 14 distinct molecules for use in humans affected by hematological or solid malignancies. In the inaugural issue of OncoImmunology, we briefly described the scientific rationale behind the use of monoclonal antibodies in cancer therapy and discussed recent, ongoing clinical studies investigating the safety and efficacy of this approach in patients. Here, we summarize the latest developments in this exciting area of clinical research, focusing on high impact studies that have been published during the last 15 months and clinical trials launched in the same period to investigate the therapeutic profile of promising, yet hitherto investigational, monoclonal antibodies. PMID:23482847

Vacchelli, Erika; Eggermont, Alexander; Galon, Jérôme; Sautès-Fridman, Catherine; Zitvogel, Laurence; Kroemer, Guido; Galluzzi, Lorenzo

2013-01-01

139

A novel pancreatic ?-cell targeting bispecific-antibody (BsAb) can prevent the development of Type 1 diabetes in NOD mice.  

PubMed

To prepare a novel Bispecific Antibody (BsAb) as a potential targeted therapy for T1D, we produced a "functionally inert" monoclonal antibody (mAb) against Glucose transporter-2 (GLUT-2) expressed on ?-cells to serve as an anchoring antibody. The therapeutic arm is an agonistic mAb against Cytotoxic T-Lymphocyte Antigen 4 (CTLA-4), a negative regulator of T-cell activation expressed on activated CD4+ T-cells. A BsAb was prepared by chemically coupling an anti-GLUT2 mAb to an agonistic anti-CTLA-4 mAb. This BsAb was able to bind to GLUT2 and CTLA-4 in vitro, and to pancreatic islets, both in vitro and in vivo. We tested the safety and efficacy of this BsAb by treating Non-Obese Diabetes (NOD) mice and found that it could delay the onset of diabetes with no apparent undesirable side effects. Thus, engagement of CTLA-4 on activated T cells from target tissue can be an effective way to treat type-1 diabetes. PMID:24792135

Bhattacharya, Palash; Fan, Jilao; Haddad, Christine; Essani, Abdul; Gopisetty, Anupama; Elshabrawy, Hatem A; Vasu, Chenthamarakshan; Prabhakar, Bellur S

2014-07-01

140

In vitro and in vivo antitumor effects of recombinant bispecific antibodies based on humanized anti-EGFR antibody.  

PubMed

We performed in vitro and in vivo experiments of the anti-epidermal growth factor receptor (EGFR) x anti-CD3 bispecific diabody (hEx3-Db) with the IgG-like bispecific antibodies (BsAbs) (hEx3-scFv-Fc and hEx3-scDb-Fc) and the anti-EGFR therapeutic antibody cetuximab to assess the effect of BsAbs on cancer growth inhibition. In vitro, efficacy of the BsAbs and cetuximab were compared by growth inhibition assays of human cell lines of bile duct (TFK-1, HuCC-T1, OCUCh-LM1), epidermoid (A431), gastric (Kato-III), colon (DLD-1, SW480), and breast (SK-BR-3, MCF-7) cancer. In vivo, in three mouse models, we evaluated the anti-tumor activity of hEx3-Db and cetuximab, assessed the effect of hEx3-Db alone, and compared the antitumor activity of hEx3-Db with the IgG-like BsAbs. In vitro, hEx3-scFv-Fc showed nearly 100% killing activity for all cell lines. Both in vitro and in vivo, hEx3-Db needed CD3-positive phenotypes to induce a growth inhibitory effect. In contrast, IgG-like BsAbs showed monotherapeutic effects in vivo by inducing antibody-dependent cellular cytotoxicity (ADCC) similar to cetuximab. However, enhancement was not observed when lymphokine-activated killer cells with the T-cell phenotype were co-injected. Results suggest that IgG-like BsAbs could not efficiently direct T lymphocytes toward tumor cells to induce ADCC due to steric hindrance on binding to CD3- and Fc-receptor-positive phenotypes. Although hEx3-scFv-Fc showed high cytotoxicity in vitro, its high molecular weight limits its usefulness. With an in vivo effect comparable to hEx3-scFv-Fc and its realistic molecular weight, hEx3-scDb-Fc shows promise as a novel recombinant therapeutic antibody and may be modified to enhance its potency by prevention of steric hindrance. PMID:21743971

Watanabe, Yasuhiro; Asano, Ryutaro; Arai, Kyoko; Shimomura, Ippei; Ogata, Hiromi; Kawaguchi, Hiroko; Hayashi, Hiroki; Ohtsuka, Hideo; Yoshida, Hiroshi; Katayose, Yu; Egawa, Shinichi; Nakanishi, Takeshi; Umetsu, Mitsuo; Yasui, Hiroshi; Ishida, Tadao; Imai, Kohzoh; Kudo, Toshio; Unno, Michiaki; Kumagai, Izumi

2011-10-01

141

Preclinical evaluation of light-activatable, bispecific anti-human CD3 antibody conjugates as anti-ovarian cancer therapeutics  

PubMed Central

The administration of anti-CD3 antibodies, either unmodified or in bispecific formats, has been shown to kill tumors. However, their activity needs to be carefully controlled. We have approached this problem by inhibiting their anti-CD3 activity until it is required. Folated anti-human CD3 antibody bispecific conjugates were therefore synthesised in which the folate portion of the conjugates remained free to bind to folate receptor (FR) expressing cancer cells, whilst their anti-CD3 activity was reversibly inhibited. On irradiation with UV-A light, the T-cell binding activity of the anti-CD3 antibody can be restored only when and where it is required, i.e., adjacent to a tumor. Conjugate bound to FR expressed on normal tissues in other parts of the body remains inactive. This report describes the preclinical in vivo testing of these conjugates in transgenic mice whose T-cells express human CD3 molecules. When the ‘cloaked’ conjugates were reactivated in the region of the primary tumor, both primary tumor growth and liver metastasis were markedly reduced. That the deliberate targeting of T-cell activity locally to the primary tumor also resulted in reduced distant metastatic growth was a key finding. Light-activatable bispecific antibody conjugates similar to those described here offer a means to control T-cell targeting with a much higher degree of specificity to tumors because they minimize potentially dangerous and unwanted side effects in non-illuminated areas. The addition of light-specific targeting to the inherent tumor specific targeting of therapeutic antibody conjugates could result in the development of safer treatments for patients.

Dessi, John

2009-01-01

142

Therapeutic monoclonal antibody for sporotrichosis  

PubMed Central

Sporotrichosis is a chronic subcutaneous mycosis that affects both humans and animals worldwide. This subcutaneous mycosis had been attributed to a single etiological agent, Sporothrix schenckii. S. schenckii exhibits considerable genetic variability, and recently, it was suggested that this taxon consists of a complex of species. Sporotrichosis is caused by traumatic inoculation of the fungus, which is a ubiquitous environmental saprophyte that can be isolated from soil and plant debris. The infection is limited to cutaneous forms, but recently, more severe clinical forms of this mycosis have been described, especially among immunocompromised individuals. The immunological mechanisms involved in the prevention and control of sporotrichosis are not well understood. Some studies suggest that cell-mediated immunity plays an important role in protecting the host against S. schenckii. In contrast, the role of the humoral immune response in protection against this fungus has not been studied in detail. In a previous study, we showed that antigens secreted by S. schenckii induced a specific humoral response in infected animals, primarily against a 70-kDa molecule, indicating a possible role of specific antibodies against this molecule in infection control. In another study by our group, we produced a mAb against a 70-kDa glycoprotein of S. schenckii to better understand the effect of the passive immunization of mice infected with S. schenckii. The results showed a significant reduction in the number of CFUs in various mice organs when the mAb was injected before or during S. schenckii infection. Similar results were observed when T-cell-deficient mice were used. The drugs of choice in the treatment of sporotrichosis require long periods, and relapses are frequently observed, primarily in immunocompromised patients. The strong protection induced by the mAb against a 70-kDa glycoprotein makes it a strong candidate as a therapeutic vaccine against sporotrichosis.

Almeida, Sandro R.

2012-01-01

143

Cancer T cell immunotherapy with bispecific antibodies and chimeric antigen receptors.  

PubMed

Solid tumors contain several different types of malignant cells. This cellular heterogeneity complicates therapy for at least two reasons. First, each subpopulation may respond differently to a given treatment. Second, cancer cells are plastic, and thus may convert from a therapy-sensitive to a therapy-resistant cell type represented by another subpopulation. Therefore, successful therapies will have to target numerous malignant cell types, not just the rapidly proliferating cells as most standard treatments do. Immunotherapies with T cells engineered to recognize cancer cells via bispecific antibodies (bsAbs) or chimeric antigen receptors (CARs) are particularly promising approaches with potential to ablate both dividing and non/slow-dividing subpopulations of cancer cells. Here, we discuss several patents associated with exceptionally effective bsAbs of the tandem single-chain variable fragment (taFv) class and untangle a part of the complex network of patents directly or indirectly related to CARs. Furthermore, we speculate on the future of bsAbs and CARs for both treatment and prevention of solid tumors such as prostate cancer. PMID:23688207

Lacher, Markus D; Provenzano, Maurizio

2013-09-01

144

Monoclonal antibodies to human estrogen receptor.  

PubMed Central

Extranuclear estrogen receptor protein (estrophilin) of MCF-7 human breast cancer cells was purified by passage of the cytosol fraction of a cell homogenate through an affinity column of estradiol linked to Sepharose by a substituted di-n-propyl sulfide bridge in the 17 alpha position. Elution with 50 micro M [3H]estradiol in 10% (vol/vol) dimethyl formamide/0.5 M sodium thiocyanate gave 40% recovery of [3H]estradiol-estrophilin showing 14% of the specific radioactivity expected for the pure complex. Serum from a Lewis rat immunized with this partially purified estradiol-receptor complex contained antiestrophilin antibodies that reacted not only with nuclear and extranuclear estradiol-receptor complexes from MCF-7 cells but also with estrophilin from rat, calf, and monkey uterus, hen oviduct, and human breast cancers. Splenic lymphocytes from the immunized rat were fused with cells of two different mouse myeloma lines (P3-X63-Ag8 and Sp2/0-Ag14) to yield hybridoma cultures, 2% of which produced antibodies to estrophilin. After cloning by limiting dilution, three hybridoma lines secreting antiestrophilin were expanded in suspension culture and as ascites tumors in athymic mice to provide substantial quantities of monoclonal antibodies that recognize mammalian but not avian estrophilin and that show different degrees of reactivity with receptor from nonprimate sources. By growing the clone from Sp2/0 in the presence of [35S]methionine, radiolabeled monoclonal IgG has been prepared. These monoclonal antibodies should prove useful in the study of estrogen receptors of human reproductive tissues, in particular for the radioimmunochemical assay and immunocytochemical localization of receptors in breast cancers.

Greene, G L; Nolan, C; Engler, J P; Jensen, E V

1980-01-01

145

Monoclonal antibodies and method for detecting dioxins and dibenzofurans  

DOEpatents

Compositions of matter are described which include five monoclonal antibodies that react with dioxins and dibenzofurans, and the five hybridomas that produce these monoclonal antibodies. In addition, a method for the use of these antibodies in a sensitive immunoassay for dioxins and dibenzofurans is given, which permits detection of these pollutants in samples at concentrations in the range of a few parts per billion.

Vanderlaan, Martin (San Ramon, CA); Stanker, Larry H. (Livermore, CA); Watkins, Bruce E. (Livermore, CA); Bailey, Nina R. (Berkley, CA)

1989-01-01

146

Labeling of monoclonal antibodies with radionuclides  

SciTech Connect

Antibodies, specifically monoclonal antibodies, are potentially very useful and powerful carriers of therapeutic agents to target tissues and diagnostic agents. The loading or charging of antibodies with agents, especially radiotracers, is reviewed here. The choice of radioisotope for immunodetection and/or immunotherapy is based on its availability, half-life, nature of the radiation emitted, and the metabolic pathways of the radionuclide in the body. Most important of all are the derivatization techniques available for labeling the antibody with the given radionuclide. Isotopes of iodine and divalent metal ions are the most commonly used radionuclides. Antibodies labeled with iodine at tyrosine residues are metabolized rapidly in vivo. This leads to the incorporation of metabolized radioactive iodine into various tissues, mainly the thyroid gland and stomach, and to the accumulation of high levels of circulating iodine in the blood, which masks tumor uptake considerably. To overcome these limitations, the use of iodohippurate as an iodine-anchoring molecule to the protein should be considered. When divalent or multivalent metal ions are used as the preferred radionuclide, bifunctional chelating reagents such as EDTA or DTPA are first coupled to the protein or antibody. These chelating molecules are attached to the protein by formation of an isopeptide linkage between the carboxylate of the chelating reagent and the amino group of the protein. Several procedures are available to generate the isopeptide linkage. When the anchoring of the chelating agent through isopeptide linkage results in the inactivation of the antibody, periodate oxidation of the carbohydrate moiety of the antibody, followed by reductive coupling of chelator, could be considered as an alternative. There is still a need for better, simpler, and more direct methods for labeling antibodies with radionuclides. 78 references.

Bhargava, K.K.; Acharya, S.A. (Albert Einstein College of Medicine-Montefiore Medical Center, Bronx, NY (USA))

1989-07-01

147

CD3 directed bispecific antibodies induce increased lymphocyte-endothelial cell interactions in vitro  

PubMed Central

Bispecific antibody (BsMAb) BIS-1 has been developed to redirect the cytolytic activity of cytotoxic T lymphocytes (CTL) to epithelial glycoprotein-2 (EGP-2) expressing tumour cells. Intravenous administration of BIS-1 F(ab?)2to carcinoma patients in a phase I/II clinical trial, caused immunomodulation as demonstrated by a rapid lymphopenia prior to a rise in plasma tumour necrosis factor-? and interferon-? levels. Yet, no lymphocyte accumulation in the tumour tissue and no anti-tumour effect could be observed. These data suggest a BsMAb-induced lymphocyte adhesion to blood vessel walls and/or generalized redistribution of the lymphocytes into tissues. In this study, we describe the effects of BIS-1 F(ab?)2binding to peripheral blood mononuclear cells (PBMC) on their capacity to interact with resting endothelial cells in vitro. Resting and pre-activated PBMC exhibited a significant increase in adhesive interaction with endothelial cells when preincubated with BIS-1 F(ab?)2, followed by an increase in transendothelial migration (tem). Binding of BIS-1 F(ab?)2to PBMC affected the expression of a number of adhesion molecules involved in lymphocyte adhesion/migration. Furthermore, PBMC preincubated with BIS-1 F(ab?)2induced the expression of endothelial cell adhesion molecules E-selectin, VCAM-1 and ICAM-1 during adhesion/tem. These phenomena were related to the CD3 recognizing antibody fragment of the BsMAb and dependent on lymphocyte–endothelial cell contact. Possibly, in patients, the BIS-1 F(ab?)2infusion induced lymphopenia is a result of generalized activation of endothelial cells, leading to the formation of a temporary sink for lymphocytes. This process may distract the lymphocytes from homing to the tumour cells, and hence prevent the occurrence of BIS-1 F(ab?)2– CTL-mediated tumour cell lysis. © 2000 Cancer Research Campaign

Molema, G; Tervaert, J W Cohen; Kroesen, B J; Helfrich, W; Meijer, D K F; Leij, L F M H de

2000-01-01

148

The Role of Monoclonal Antibodies in the Management of Leukemia  

PubMed Central

This article will review the monoclonal antibodies more commonly used in leukemias. In the last three decades, scientists have made considerable progress understanding the structure and the functions of various surface antigens, such as CD20, CD33. The introduction of rituximab, an anti CD20 monoclonal antibody, had a great impact in the treatment of lymphoproliferative disorders. Gemtuzumab, an anti CD 33 conjugated monoclonal antibody has activity in acute mylegenous leukemia (AML). As this field is undergoing a rapid growth, the years will see an increasing use of monoclonal antibodies in hematological malignancies.

Al-Ameri, Ali; Cherry, Mohamad; Al-Kali, Aref; Ferrajoli, Alessandra

2010-01-01

149

Comparison of physical chemical properties of llama V HH antibody fragments and mouse monoclonal antibodies  

Microsoft Academic Search

Antigen specific llama VHH antibody fragments were compared to antigen specific mouse monoclonal antibodies with respect to specificity, affinity and stability. The llama VHH antibody fragments and the mouse monoclonal antibodies investigated were shown to be highly specific for the protein antigen hCG or the hapten antigen RR-6. The affinity of the interaction between monovalent llama VHH antibody fragments and

R. H. J. van der Linden; L. G. J. Frenken; B. de Geus; M. M. Harmsen; R. C. Ruuls; W. Stok; L. de Ron; S. Wilson; P. Davis; C. T. Verrips

1999-01-01

150

Phylogenetic study of transcortin using monoclonal antibodies.  

PubMed

We produced monoclonal antibodies that recognise three distinct epitopes of human transcortin. These epitopes are present on transcortin of humans with normal and altered transcortin levels, as well as on a variant with lower affinity for cortisol. One epitope is present on transcortin of Old World Monkeys and apes, the others are only present on transcortin of apes. The epitopes are not present on transcortin of other species. These results indicate that human transcortin contains a highly evolved and a more conserved part. PMID:2428359

Faict, D; De Moor, P

1986-08-14

151

Monoclonal antibodies to oxytocin: production and characterization.  

PubMed

To induce a good immune response to oxytocin (OT) we developed a two-step technique to conjugate OT to thyroglobulin (TG) using glutaraldehyde. We obtained 30 hybridomas recognizing OT-ovalbumin conjugates and 16 stable lines. Three monoclonal antibodies were selected for further characterization. One of them (O13) was found very specific for OT using three different techniques (enzyme-linked immunosorbent assay, radioimmunoassay and immunohistochemistry); it is directed to the C-terminal tripeptide. The other two probably recognize tyrosine containing epitope(s) also shared by vasopressin and other related nonapeptides. PMID:1995653

Burgeon, E; Chapleur, M; Schoenen, J; Remichius, D; Legros, J J; Geenen, V; Robert, F

1991-03-01

152

Further characterization of cooperative interactions of monoclonal antibodies.  

PubMed

We reported previously that mixtures of some monoclonal antibodies directed against the glycoprotein hormone human chorionic gonadotropin (hCG) had a higher affinity for the antigen than either monoclonal antibody separately. The synergistic interaction could no longer be detected when one of the antibodies was replaced with its F(ab) fragment. This cooperative interaction has now been further characterized. One-half of 10 possible pairs prepared from five IgG1 monoclonal antibodies against hCG result in a synergistic interaction. The addition of an IgG2b monoclonal antibody to one of the IgG1 monoclonal antibodies also induces a cooperative interaction, which shows that the effect is not subclass restricted. Cooperative interactions between antibodies are also not restricted to solution conditions; adsorption of one antibody to a solid support appears to increase the cooperative effect. Indeed, one pair of antibodies that failed to bind hCG synergistically in solution did so when one antibody was bound to a solid surface. The liquid phase antibody also has an effect on the specificity of the solid phase antibody. The sensitivity of the solid phase assay system has enabled us to develop a rapid method of determining if two monoclonal antibodies can bind to an antigen simultaneously. A quantitative theoretical model has been devised that successfully predicts the cooperative behavior observed between antibodies and should be useful in devising conditions that result in sensitive solid phase radioimmunoassays. PMID:6619545

Ehrlich, P H; Moyle, W R; Moustafa, Z A

1983-10-01

153

Development of Monoclonal Antibodies to Trypanosoma brucei rhodesiense Antigens.  

National Technical Information Service (NTIS)

This report, covering work performed from 1 September 1979-28 February 1982,describes methdology of tissue culture, hybridization procedures, immunization schedules and antibody assays developed to produce monoclonal antibodies to variant specific surface...

G. H. Campbell

1982-01-01

154

Development of Monoclonal Antibodies to Trypanosoma brucei rhodesiense Antigens.  

National Technical Information Service (NTIS)

This report, describes methodology of tissue culture, hybridization procedures, immunization schedules and antibody assays developed to produce monoclonal antibodies to variant specific surface antigens of Trypansoma b. rhodesiense organisims of the Walte...

G. H. Campbell J. Giorgi

1980-01-01

155

NCI Requests Cancer Targets for Monoclonal Antibody Production and Characterization  

Cancer.gov

In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

156

Monoclonal antibodies to Pseudomonas aeruginosa ferripyochelin-binding protein.  

PubMed Central

Hybridomas secreting specific monoclonal antibodies against the Pseudomonas aeruginosa ferripyochelin-binding protein (FBP) were isolated. These monoclonal antibodies reacted with FBP in immunoblots of outer membrane preparations from all serotypes of P. aeruginosa. Two of the monoclonal antibodies also reacted with FBP in strains of P. putida, P. fluorescens, and P. stutzeri. These antibodies did not react with outer membranes of P. cepacia, "P. multivorans," P. maltophilia, or other gram-negative organisms. The monoclonal antibodies were opsonophagocytic and blocked the binding of [59Fe]ferripyochelin to isolated outer membranes of strain PAO. By indirect immunofluorescence techniques, the monoclonal antibodies were used to demonstrate that FBP is present on the cell surface of P. aeruginosa cells grown in low-iron but not high-iron medium. These observations were confirmed by using 125I in surface-labeling techniques. Images

Sokol, P A; Woods, D E

1986-01-01

157

Virotherapy, gene transfer and immunostimulatory monoclonal antibodies  

PubMed Central

Malignant cells are susceptible to viral infection and consequent cell death. Virus-induced cell death is endowed with features that are known to stimulate innate and adaptive immune responses. Thus danger signals emitted by cells succumbing to viral infection as well as viral nucleic acids are detected by specific receptors, and tumor cell antigens can be routed to professional antigen-presenting cells. The anticancer immune response triggered by viral infection is frequently insufficient to eradicate malignancy but may be further amplified. For this purpose, transgenes encoding cytokines as co-stimulatory molecules can be genetically engineered into viral vectors. Alternatively, or in addition, it is possible to use monoclonal antibodies that either block inhibitory receptors of immune effector cells, or act as agonists for co-stimulatory receptors. Combined strategies are based on the ignition of a local immune response at the malignant site plus systemic immune boosting. We have recently reported examples of this approach involving the Vaccinia virus or Semliki Forest virus, interleukin-12 and anti-CD137 monoclonal antibodies.

Quetglas, Jose I.; John, Liza B.; Kershaw, Michael H.; Alvarez-Vallina, Luis; Melero, Ignacio; Darcy, Phillip K.; Smerdou, Cristian

2012-01-01

158

Trifunctional bispecific antibodies induce tumor-specific T cells and elicit a vaccination effect.  

PubMed

A major goal of tumor immunotherapy is the induction of long-lasting systemic T-cell immunity. Bispecific antibodies (bsAbs) that lack the immunoglobulin Fc region confer T-cell-mediated killing of tumor cells but do not induce long-term memory. In contrast, trifunctional bsAbs comprise an appropriate Fc region and, therefore, not only recruit T cells but also accessory cells that bear activating Fc? receptors (Fc?R), providing additional T-cell-activating signals and securing presentation of tumor-derived antigens to T cells. In this study, we show that trifunctional bsAbs induce a polyvalent T-cell response and, therefore, a vaccination effect. Mice were treated with melanoma cells and with a trifunctional bsAb directed against the melanoma target antigen ganglioside GD2 in addition to murine CD3. The trifunctional bsAb activated dendritic cells and induced a systemic immune response that was not replicated by treatment with the F(ab')2-counterpart lacking the Fc region. Restimulation of spleen and lymph node cells in vitro yielded T-cell lines that specifically produced interferon-? in response to tumor. In addition, trifunctional bsAb-induced T cells recognized various specific peptides derived from melanoma-associated antigens. Moreover, these polyvalent responses proved to be tumor-suppressive and could not be induced by the corresponding bsF(ab')2-fragment. Taken together, our findings provide preclinical proof of concept that trifunctional bsAbs can induce tumor-specific T cells with defined antigen specificity. PMID:22745368

Eissler, Nina; Ruf, Peter; Mysliwietz, Josef; Lindhofer, Horst; Mocikat, Ralph

2012-08-15

159

Functional comparison of single-chain and two-chain anti-CD3-based bispecific antibodies in gene immunotherapy applications  

PubMed Central

Gene therapy to achieve in vivo secretion of recombinant anti-CD3 x anti-tumor bispecific antibodies in cancer patients is being explored as a strategy to counterbalance rapid renal elimination, thereby sustaining levels of bispecific antibodies in the therapeutic range. Here, we performed a comparative analysis between single- and two-chain configurations for anti-CD3 x anti-CEA (carcinoembryonic antigen) bispecific antibodies secreted by genetically-modified human cells. We demonstrate that tandem single-chain variable fragment (scFv) antibodies and two-chain diabodies are expressed as soluble secreted proteins with similar yields. However, we found significant differences in their biological functionality (i.e., antigen binding) and in their ability to induce non-specific T cell activation. Whereas single-chain tandem scFvs induced human T cell activation and proliferation in an antigen-independent manner, secreted two-chain diabodies exerted almost no proliferative stimulus when human T cells were cultured alone or in co-cultures with CEA negative cells. Thus, our data suggest that two-chain diabodies are preferable to single-chain tandem scFvs for immunotherapeutic strategies comprising in vivo secretion of bispecific antibodies aiming to recruit and activate anticancer specific lymphocytic effector T cells.

Compte, Marta; Alvarez-Cienfuegos, Ana; Nunez-Prado, Natalia; Sainz-Pastor, Noelia; Blanco-Toribio, Ana; Pescador, Nuria; Sanz, Laura; Alvarez-Vallina, Luis

2014-01-01

160

DETECTION OF ROTAVIRUS IN HUMAN STOOLS BY USING MONOCLONAL ANTIBODY  

EPA Science Inventory

A monoclonal antibody, 3F7, which reacts with the common rotavirus antigen on the sixth viral gene product was prepared. It was used in a direct monoclonal antibody radioimmunoassay (RIA) as a diagnostic reagent for detection in 3.5 hours of rotavirus in human pediatric stool spe...

161

Monoclonal antibodies to detect human tumours: an experimental approach  

Microsoft Academic Search

The use of monoclonal antibodies which can be raised to antigens of choice offers a selective and specific approach for the detection of tumours both in vivo and at a cellular level in biopsy specimens. We demonstrate that a monoclonal antibody raised to human teratoma will localise in a teratoma, growing as a xenograft in immune-suppressed mice.

V Moshakis; R McIlhinney; D Raghavan; A M Neville

1981-01-01

162

Emerging Therapies: Spectrum of Applications of Monoclonal Antibody Therapy  

Microsoft Academic Search

This article focuses on the recent dramatic ad- vances in the applications of monoclonal antibody therapy to hematopoietic and neoplastic disease. The increase in the understanding of the role of growth factors and their receptors in the pathogen- esis of malignancy and other undesirable hemato- logical events taken in conjunction with the ability to produce humanized chimeric monoclonal antibodies to

Thomas A. Waldmann; Ronald Levy; Barry S. Coller

163

Hierarchical Clustering of Monoclonal Antibody Reactivity Patterns in Nonhuman Species  

PubMed Central

Monoclonal antibodies are an important resource for defining molecular expression and probing molecular function. The characterization of monoclonal antibody reactivity patterns, however, can be costly and inefficient in nonhuman experimental systems. To develop a computational approach to the pattern analysis of monoclonal antibody reactivity, we analyzed a panel of 128 monoclonal antibodies recognizing sheep antigens. Quantitative single parameter flow cytometry histograms were obtained from five cell types isolated from normal animals. The resulting 640 histograms were smoothed using a Gaussian kernel over a range of bandwidths. Histogram features were selected by SiZer—an analytic tool that identifies statistically significant features. The extracted histogram features were compared and grouped using hierarchical clustering. The validity of the clustering was indicated by the accurate pairing of externally verified molecular reactivity. We conclude that our computational algorithm is a potentially useful tool for both monoclonal antibody classification and molecular taxonomy in nonhuman experimental systems.

Pratt, Juan Pablo; Zeng, Qing; Ravnic, Dino; Huss, Harold; Rawn, James; Mentzer, Steven J.

2010-01-01

164

Preparation and characterization of monoclonal antibodies to swine lymphocyte antigens  

SciTech Connect

A panel of hybridoma lines were produced by the fusion of Sp2/0 myeloma cells and spleen cells from mice immunized with pig peripheral blood lymphocytes. Thirty-three stable hybridomas were produced which secreted monoclonal antibodies that reacted with pig lymphocytes, as determined by an ELISA screening procedure. These monoclonal antibodies were characterized by a complement-mediated cytotoxicity test and by flow cytometric analysis. The molecular weights of the antigens recognized by the monoclonal antibodies were determined by immunoprecipitation of /sup 125/I surface labeled lymphocytes, followed by SDS-PAGE. One monoclonal antibody, 7-34-1 (IgG2a), which reacted to all peripheral blood lymphocytes, precipitated a MHC class I molecule composed of a 50 kd heavy chain and a 12 kd light chain (..beta../sub 2/ microglobulin). This monoclonal antibody may prove to be an important reagent for the detection of SLA antigens on pig cells.

Lie, W.R.; Rothschild, M.F.; Warner, C.M.

1986-03-05

165

The immunodiagnosis of lung cancer with monoclonal antibodies.  

PubMed

Lung cancer is responsible for much suffering and death worldwide. The only hope for cure is therapy applied in an early phase, and all methods of diagnosis should be aimed at this goal. This paper reviews the development of the use of monoclonal antibodies in the diagnosis of lung cancer. Relevant data since the publication of the technology of producing monoclonal antibodies in 1975 to the present are summarized. The authors evaluate the progress of the immunodiagnosis of lung cancer by monoclonal antibodies from pleural effusion, bone marrow, sputum, bronchial lavage, and bronchial brush (immunocytochemistry). They collect recent data on the immunohistochemistry of biopsy materials and of removed tissues. They evaluate radioimmuno-imaging (radioimmuno-scintigraphy) and immuno-PET as in vivo macroscopic diagnostic methods of lung cancer by monoclonal antibodies as well as the help monoclonal antibodies provide in radioimmuno-guided surgery or immunoimage-guided, focally ablative therapy of this disease. PMID:16127376

Egri, Gábor; Takáts, Alajos

2005-09-01

166

Development of syngeneic monoclonal anti-idiotype antibodies to mouse monoclonal anti-asialoglycoprotein receptor antibody.  

PubMed

Anti-idiotype antibodies (Ab2) play an important role in the homeostasis of immune responses and are related to the development and the disease activity of certain autoimmune diseases. The asialoglycoprotein receptor (ASGPR) is considered one of the target antigens in the pathogenesis of autoimmune chronic active hepatitis (AIH). We previously developed a mouse monoclonal antibody (clone 8D7) which recognizes rat and human ASGPR. In this study, to help investigate the anti-ASGPR antibody-anti-idiotype antibody network in patients with AIH, we developed a syngeneic mouse monoclonal Ab2 to the 8D7 anti-ASGPR antibody (Ab1). One clone, designated as 3C8, tested positive for specific reactivity to 8D7-Ab1 and did not bind to other irrelevant immunoglobulins. By competitive inhibition assays, the binding of 8D7-Ab1 to liver membrane extracts, i.e., the crude antigen preparation, was inhibited by 3C8-Ab2 in a dose-dependent manner, and the binding of 8D7-Ab1 to 3C8-Ab2 was inhibited by the liver membrane extracts. In the immunohistochemical analysis, 3C8-Ab2 blocked the specific staining of sinusoidal margins of rat hepatocytes by 8D7-Ab1. These results suggest that 3C8 anti-idiotype antibody recognizes the specific idiotypic determinants within the antigen-binding site of 8D7-Ab1. PMID:12108584

Hirai, Michio; Mizuno, Motowo; Morisue, Yoshiko; Yoshioka, Masao; Shimada, Morizou; Nasu, Junichirou; Okada, Hiroyuki; Shimomura, Hiroyuki; Yamamoto, Kazuhide; Tsuji, Takao

2002-06-01

167

Monoclonal antibodies against Xenopus greatwall kinase.  

PubMed

Mitosis is known to be regulated by protein kinases, including MPF, Plk1, Aurora kinases, and so on, which become active in M-phase and phosphorylate a wide range of substrates to control multiple aspects of mitotic entry, progression, and exit. Mechanistic investigations of these kinases not only provide key insights into cell cycle regulation, but also hold great promise for cancer therapy. Recent studies, largely in Xenopus, characterized a new mitotic kinase named Greatwall (Gwl) that plays essential roles in both mitotic entry and maintenance. In this study, we generated a panel of mouse monoclonal antibodies (MAbs) specific for Xenopus Gwl and characterized these antibodies for their utility in immunoblotting, immunoprecipitation, and immunodepletion in Xenopus egg extracts. Importantly, we generated an MAb that is capable of neutralizing endogenous Gwl. The addition of this antibody into M-phase extracts results in loss of mitotic phosphorylation of Gwl, Plk1, and Cdk1 substrates. These results illustrate a new tool to study loss-of-function of Gwl, and support its essential role in mitosis. Finally, we demonstrated the usefulness of the MAb against human Gwl/MASTL. PMID:22008075

Wang, Ling; Fisher, Laura A; Wahl, James K; Peng, Aimin

2011-10-01

168

T-cell activation and B-cell depletion in chimpanzees treated with a bispecific anti-CD19\\/anti-CD3 single-chain antibody construct  

Microsoft Academic Search

BscCD19xCD3 is a bispecific single-chain antibody construct with exceptional cytotoxic potency in vitro and in vivo. Here, we have investigated the biological activity of bscCD19xCD3 in chimpanzee, the only animal species identified in which bscCD19xCD3 showed bispecific binding, redirected B-cell lysis and cytokine production comparable to human cells. Pharmacokinetic analysis following 2-h intravenous infusion of 0.06, 0.1 or 0.12 ?g\\/kg of

Bernd Schlereth; Cornelia Quadt; Torsten Dreier; Peter Kufer; Grit Lorenczewski; Nadja Prang; Christian Brandl; Sandra Lippold; Kathy Cobb; Kathleen Brasky; Eugen Leo; Ralf Bargou; Krishna Murthy; Patrick A. Baeuerle

2006-01-01

169

The birth pangs of monoclonal antibody therapeutics  

PubMed Central

This paper examines the development and termination of nebacumab (Centoxin®), a human IgM monoclonal antibody (mAb) drug frequently cited as one of the notable failures of the early biopharmaceutical industry. The non-approval of Centoxin in the United States in 1992 generated major concerns at the time about the future viability of any mAb therapeutics. For Centocor, the biotechnology company that developed Centoxin, the drug posed formidable challenges in terms of safety, clinical efficacy, patient selection, the overall economic costs of health care, as well as financial backing. Indeed, Centocor's development of the drug brought it to the brink of bankruptcy. This article shows how many of the experiences learned with Centoxin paved the way for the current successes in therapeutic mAb development.

2012-01-01

170

Monoclonal antibodies in treatment of multiple sclerosis.  

PubMed

Monoclonal antibodies (mAbs) are used as therapeutics in a number of disciplines in medicine, such as oncology, rheumatology, gastroenterology, dermatology and transplant rejection prevention. Since the introduction and reintroduction of the anti-alpha4-integrin mAb natalizumab in 2004 and 2006, mAbs have gained relevance in the treatment of multiple sclerosis (MS). At present, numerous mAbs have been tested in clinical trials in relapsing-remitting MS, and in progressive forms of MS. One of the agents that might soon be approved for very active forms of relapsing-remitting MS is alemtuzumab, a humanized mAb against CD52. This review provides insights into clinical studies with the mAbs natalizumab, alemtuzumab, daclizumab, rituximab, ocrelizumab and ofatumumab. PMID:24001305

Rommer, P S; Dudesek, A; Stüve, O; Zettl, U K

2014-03-01

171

Monoclonal antibody dose determination and biodistribution into solid tumors.  

PubMed

Monoclonal antibodies are increasingly being used as protein therapeutics for cancer. They offer very specific binding to target molecules on the surface of cancer cells, relatively few side effects and predictable pharmacokinetics. Tumor shrinkage is seen in some patients, and an incremental improvement in survival occurs in the group. However, due to their large size and consequent slow diffusion, antibody penetration deep into tumors may be inhomogeneous. Even if only a few cells, deep in tumors, escape therapy, they can regrow and lead to clinical relapse, limiting the significant potential of monoclonal antibody therapy. This leads to questions about optimal dosing for monoclonal antibodies. Methods to determine monoclonal antibody dose include maximum-tolerated dose studies, pharmacokinetically and pharmacodynamically guided dosing, randomized dose-ranging studies, imaging of antibody biodistribution and competitive-binding studies. Limitations of these methods, and future directions to possibly overcome these limitations will be discussed. PMID:22834004

Beckman, Robert A; von Roemeling, Reinhard; Scott, Andrew M

2011-03-01

172

Regulatory T cells are redirected to kill glioblastoma by an EGFRvIII-targeted bispecific antibody  

PubMed Central

Regulatory T cells (Tregs) play a central role in in tumor escape from immunosurveillance. We report that a bispecific T-cell engager (BiTE) targeting a mutated form of the epidermal growth factor receptor, i.e., EGFRvIII, potently redirects Tregs to kill glioblastoma through the granzyme-perforin pathway.

Choi, Bryan D; Gedeon, Patrick C; Sanchez-Perez, Luis; Bigner, Darell D; Sampson, John H

2013-01-01

173

The use of monoclonal antibodies for treatment of autoimmune disease  

Microsoft Academic Search

Over the past decade monoclonal antibodies have been successfully employed in a number of animal models of autoimmune disease. We have used antibodies to the class II gene products of the major histocompatibility complex, the CD4 molecule on helper T cells, and the T-cell receptor. Monoclonal anti-class II antibodies have been administered to treat paralytic disease in the animal model

Lawrence Steinman

1990-01-01

174

Monoclonal antibodies to human thioredoxin reductase.  

PubMed

The thioredoxin system consisting of thioredoxin (Trx), thioredoxin reductase (TrxR), and NADPH is an electron donor for ribonucleotide reductase but has also been implicated in other cellular events, including secretion, growth promotion, regulation of transcription factors, protection against oxidative stress, and apoptosis. Mammalian TrxR is a dimeric flavoprotein with 58 kDa subunits each with a catalytically active selenocysteine residue. To study the function and expression of TrxR, we have produced and characterized, for the first time, monoclonal antibodies against human TrxR. Native placenta TrxR was used for immunization of BALB/c mice, followed by hybridization, cloning, and establishment of hybridomas producing specific antibodies against human TrxR. Three clones of IgG1, kappa subclass, termed anti-TrxR1, anti-TrxR2, and anti-TrxR3, were studied in detail. The isoelectric points (pIs) of the mAbs were 6.5, 6.0, and 6.5, respectively. The affinities (Ka) of the mAbs were 2 x 10(8) M-1. Inhibition ELISA using biotin-labeled versus nonconjugated mAb IgG revealed that all three mAbs recognized one immunodominant epitope. Western blot analysis showed that the antibodies specifically bound to a 58 kDa protein, representing the subunit of TrxR. A Trx-dependent insulin reduction assay was used for analysis of enzymatic activity and the antibodies neutralized the reductase activity. PMID:9705836

Söderberg, A; Sahaf, B; Holmgren, A; Rosén, A

1998-08-10

175

Superior antitumor activity of a novel bispecific antibody cotargeting human epidermal growth factor receptor 2 and type I insulin-like growth factor receptor.  

PubMed

The humanized anti-HER2 monoclonal antibody (mAb) trastuzumab (Herceptin; Genentech) effectively inhibits human epidermal growth factor receptor 2 (HER2)-positive breast tumors. However, many patients responding to treatment often develop resistance. Cross-talk between type I insulin-like growth factor receptor (IGF-IR) and HER2 and elevated IGF-IR signaling have been implicated in tumor cell resistance to trastuzumab therapy. Previously, we reported that the anti-IGF-IR mAb m590 inhibits proliferation and migration of breast cancer MCF-7 cells in vitro. Here, we generated a "knobs-into-holes" bispecific antibody (Bi-Ab) against HER2 and IGF-IR by engineering trastuzumab and m590. We compared the effects of Bi-Ab treatment in vitro and in SKOV-3 HER2- and IGF-IR-overexpressing cancer xenograft mouse model with those of m590 and trastuzumab treatment alone or in combination. Bi-Ab effectively inhibited proliferation of HER2- and IGF-IR-overexpressing ovarian cancer SKOV-3 cells in vitro by ablating receptor phosphorylation and downstream PI3K/Akt and mitogen-activated protein kinase signaling. Bi-Ab more effectively inhibited cancer growth in SKOV-3 HER2- and IGF-IR-overexpressing cancer xenograft mouse model than m590 and trastuzumab alone or in combination. Mice bearing SKOV-3 HER2- and IGF-IR-overexpressing xenografts showed extensive and sustainable tumor regression when treated with Bi-Ab. Our results suggest that Bi-Ab has superior antitumor activity compared with monospecific antibodies, and cotargeting HER2 and IGF-IR may be clinically beneficial in minimizing the acquired resistance to trastuzumab therapy. PMID:24227890

Chen, Chao; Zhang, Yanyu; Zhang, Yu; Li, Jingjing; Tsao, Sai Wah; Zhang, Mei-Yun

2014-01-01

176

Serological analysis of serogroup icterohaemorrhagiae using monoclonal antibodies.  

PubMed

Eighteen serovars (19 strains) of serogroup Icterohaemorrhagiae were serologically analyzed using 18 monoclonal antibodies against serovar copenhageni Shiromizu, M20 and serovar icterohaemorrhagiae RGA strains. The reaction patterns of the serovars against these monoclonal antibodies were different. According to these results, we divided the serovars, except for serovar tonkini, into the following three subgroups: Subgroup 1 reacted to many monoclonal antibodies including serovars icterohaemorrhagiae, copenhageni, hualien, monymusk, mankarso, and budapest. Subgroup 2 fell between subgroups 1 and 3 including serovars dakota, naam, bogvere, birkini, smithi, ndambari, gem, ndahambukuje and mwogolo. Subgroup 3 reacted to only a few monoclonal antibodies: serovars weaveri and sarmin. Serovar tonkini did not react to any of the monoclonal antibodies used. There is a possibility that serovar tonkini does not belong to serogroup Icterohaemorrhagiae. Further studies on the serological reactions of each strain revealed that it was impossible to distinguish the RGA strain from the serovar hualien LT11-31 strain, indicating that they may be identical. It was also observed that serovar copenhageni and monymusk seemed to be closely related. Serovars birkini and smithi, and serovars ndambari and gem were alike in their serological reactivities. Among the 18 monoclonal antibodies, RGAMA-1 was a unique antibody which reacted only to serovar icterohaemorrhagiae and serovar hualien, indicating that it must be the serovar icterohaemorrhagiae specific antibody. On the other hand, SHIRMA-2, 5, 6 reacted to all the serovars except for serovars weaveri, sarmin, and tonkini. These antibodies exhibited a broad reaction spectrum. PMID:3831722

Kobayashi, Y; Tamai, T; Sada, E

1985-01-01

177

Clinical laboratory applications of monoclonal antibodies.  

PubMed Central

Monoclonal antibody (MAb) technology is well recognized as a significant development for producing specific serologic reagents to a wide variety of antigens in unlimited amounts. These reagents have provided the means for developing a number of highly specific and reproducible immunological assays for rapid and accurate diagnosis of an extensive list of diseases, including infectious diseases. The impact that MAbs have had in characterizing infectious disease pathogens, as well as their current and future applications for use in clinical microbiology laboratories, is reviewed. In addition, the advantages (and disadvantages) of the use of MAbs in a number of immunoassays, such as particle agglutination, radioimmunoassays, enzyme-linked immunosorbent assays, immunofluorescent-antibody assays, and immunohistology, are explored, including the use of these reagents in novel test system assays. Also, nucleic acid probe technology is compared with the use of MAbs from the perspective of their respective applications in the diagnosis of infectious disease agents. There is no question that hybridoma technology has the potential to alter significantly the methods currently used in most clinical microbiology laboratories.

Payne, W J; Marshall, D L; Shockley, R K; Martin, W J

1988-01-01

178

Rat monoclonal antibodies against Aspergillus galactomannan.  

PubMed Central

Monoclonal antibodies (MAbs) against Aspergillus fumigatus galactomannan were produced in rats. Seven of them, EB-A1 through EB-A7, were characterized in more detail. They were all immunoglobulin M antibodies, reacting in an indirect enzyme-linked immunosorbent assay with purified A. fumigatus galactomannan, with avidity constants of between 2 x 10(9) and 5 x 10(9)/M. Enzyme-linked immunosorbent assay inhibition experiments with modified galactomannan and synthetic oligomers of beta (1----5)galactofuranose demonstrated that the MAbs bound to an epitope located on the beta(1----5)galactofuranose-containing side chains of the galactomannan molecule. An identical or similar epitope also seemed to be present in other fungi. Immunofluorescence and immunoelectron microscopy experiments with EB-A2 revealed the presence of the antigen in the fungal wall and inside the cell. Immunoblotting experiments demonstrated that the epitope recognized by the MAbs was a common oligosaccharide moiety of a wide range of intracellular and extracellular glycoproteins in A. fumigatus. The characteristics of the MAbs justify their use in the diagnosis of invasive aspergillosis by antigen detection. Images

Stynen, D; Sarfati, J; Goris, A; Prevost, M C; Lesourd, M; Kamphuis, H; Darras, V; Latge, J P

1992-01-01

179

Monoclonal antibody specific for a pigmentation associated antigen  

SciTech Connect

Monoclonal antibody TA99, which specifically binds to a pigmentation associated antigen present on melanoma cells is described. Additionally, the hybridoma cell line deposited with the ATCC under Accession Number HB 8704 from which the antibody is derived, as well as methods for using the antibody are described.

Thomson, T.M.; Mattes, M.J.; Old, L.J.; Lloyd, K.O

1989-01-17

180

Functional Studies of Willebrand Factor Using Monoclonal Antibodies  

Microsoft Academic Search

Previously described monoclonal antibodies to porcine Wil- lebrand factor were used to study various functional assays of Willebrand factor. These antibodies comprise at least five separate specificities as determined by differential reactivity in three distinct binding assays. The antibodies were tested for their effect on the in vivo bleeding time. in vitro bleeding time. and ristocetin cofactor activity. The titers

E. J. W. Bowie; D. N. Fass; J. A. Katzmann

1983-01-01

181

Monoclonal antibody directed against RNA polymerase II of Drosophila melanogaster  

Microsoft Academic Search

Monoclonal antibodies were raised against purified RNA polymerase II (or B) from Drosophila melanogaster. The antibody produced by one hybridoma cell clone was found to be directed against the two large subunits of the enzyme. The absence of antibodies directed against proteins possibly contaminating the antigens used for immunization allowed us to identify RNA polymerase unequivocally in interbands and puffs

Angela Krfimer; Regina Haars; Rainer Kabisch; Hans Will; Friedlinde A. Bautz; Ekkehard K. F. Bautz

1980-01-01

182

Human and Primate MonoClonal Antibodies for in Vivo Therapy  

Microsoft Academic Search

Human monoclonal antibodies, owing to their decreased immunogenicity, are expected to be an improvement over mouse monoclonal antibodies for in vivo therapy. Human and primate monoclonal antibodies are best produced with a human x mouse heteromyeloma. Several human chromo- somes are stable in the human x (human x mouse) hybrids. Chimpanzee anti-digoxin monoclonal antibodies were pre- pared and characterized. Because

Paul H. EhrIlch; Zelnab A Moustafa; James C. JustIce; K. Ellsabeth Harfeldt; Inder IC Gadi; Leonard J. Sclorra; Frank P. Uhl

1988-01-01

183

Monoclonal antibodies in animal production; their use in diagnostics and passive immunization  

Microsoft Academic Search

One of the landmarks in immunology was the invention and development of monoclonal antibody-secreting hybridomas by Milstein and his coworkers. The enormous promise of monoclonal antibody technology, which became apparent soon after its discovery, may explain the unusual speed with which monoclonal antibodies have been applied to biological and medical sciences.In animal production monoclonal antibodies are increasingly finding application in

P. Booman

1989-01-01

184

The Fc-region of a new class of intact bispecific antibody mediates activation of accessory cells and NK cells and induces direct phagocytosis of tumour cells  

PubMed Central

Bispecific antibodies (bsAb) are considered as promising tools for the elimination of disseminated tumour cells in a minimal residual disease situation. The bsAb-mediated recruitment of an immune effector cell in close vicinity of a tumour cell is thought to induce an antitumoural immune response. However, classical bispecific molecules activate only a single class of immune effector cell that may not yield optimal immune responses. We therefore constructed an intact bispecific antibody, BiUII (anti-CD3 × anti-EpCAM), that not only recognizes tumour cells and T lymphocytes with its two binding arms, but also binds and activates Fc?-receptor positive accessory cells through its Fc-region. We have demonstrated recently that activated accessory cells contribute to the bsAb-induced antitumoural activity. We now analyse this stimulation in more detail and demonstrate here the BiUll-induced upregulation of activation markers like CD83 and CD95 on accessory cells and the induction of neopterin and biopterin synthesis. Experiments with pure cell subpopulations revealed binding of BiUll to CD64+ accessory cells and CD16+ NK cells, but not to CD32+ B lymphocytes. We provide further evidence for the importance of the Fc-region in that this bispecific molecule stimulates Fc?-R-positive accessory cells to eliminate tumour cells in vitro by direct phagocytosis. © 2000 Cancer Research Campaign

Zeidler, R; Mysliwietz, J; Csanady, M; Walz, A; Ziegler, I; Schmitt, B; Wollenberg, B; Lindhofer, H

2000-01-01

185

Anti-Mesothelin Monoclonal Antibodies for the Treatment of Cancer  

Cancer.gov

The National Cancer Institute, Laboratory of Molecular Biology seeks parties interested in collaborative research to further co-develop monoclonal antibodies for the treatment of mesothelin-expressing cancers.

186

Human and Improved Murine Monoclonal Antibodies Against CD22  

Cancer.gov

The National Cancer Institute's Nanobiology Program is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize human monoclonal antibodies expressed in types of lymphoma.

187

Production and Characterization of Monoclonal Antibodies to Bacteroides gingivalis.  

National Technical Information Service (NTIS)

The rapid detection of Bacteroides gingivalis by immunological methods using monoclonal antibodies could greatly improve the diagnosis and prognosis of severe periodontal disease in adults. In this study, three distinct hybridomas were produced which secr...

L. G. Simonson B. R. Merrell R. F. Rouse I. L. Shklair

1986-01-01

188

Internal radiation dosimetry for clinical testing of radiolabeled monoclonal antibodies.  

National Technical Information Service (NTIS)

In gauging the efficacy of radiolabeled monoclonal antibodies in cancer treatment, it is important to know the amount of radiation energy absorbed by tumors and normal tissue per unit administered activity. This paper describes methods for estimating abso...

D. R. Fisher J. S. Durham T. E. Hui R. L. Hill

1990-01-01

189

Monoclonal Antibodies to Soybean Kunitz Trypsin Inhibitor and Immunoassay Methods.  

National Technical Information Service (NTIS)

The invention relates to and has among its objects the provision of hybridomas that produce and secrete monoclonal antibodies which are specific for soybean Kunitz trypsin inhibitor, and to immunoassay methods for the determination of active Kunitz trypsi...

D. L. Brandon A. H. Bates M. Friedman

1987-01-01

190

Reshaped Human Monoclonal Antibodies for Therapy and Passive Immunization.  

National Technical Information Service (NTIS)

The purpose of the project is to apply reshaping technology for the humanization of monoclonal antibodies specific for junin and vaccinia viruses. The cloning of immunoglobulin variable region heavy (VH) and light (VK) chain genes was achieved through syn...

W. J. Harris

1990-01-01

191

Monoclonal Antibodies: Basic and Medical Applications for Pathogenic Fungi.  

National Technical Information Service (NTIS)

The successful application of monoclonal antibody technology for diagnosing and treating deep fungal infections requires the solution of 3 interrelated problems: (1) characterization of fungal surface antigens associated with IgG as well as IgM responses,...

D. J. Gennevois J. W. Hoffman H. B. Levine A. E. Kara

1984-01-01

192

Monoclonal Antibodies to Epizootic Hemorrhagic Disease Virus Antigen.  

National Technical Information Service (NTIS)

A method for preparing hybrid cell lines (hybridomas) which secrete monoclonal antibody which is group-specific to epizootic hemorrhagic disease virus antigen and which does not react to antigenically-related bluetongue virus antigen is disclosed. The ant...

M. M. Jochim S. C. Jones

1985-01-01

193

DEVELOPMENT OF MONOCLONAL ANTIBODIES AGAINST FATHEAD MINNOW (PIMEPHALES PROMELAS) VITELLOGENIN  

EPA Science Inventory

We have obtained a panel of monoclonal antibodies directed against fathead minnow vitellogenin (Vtg) for use in sensitive ELISAs to quantify the response of exposure in vivo to estrogen or estrogen mimics....

194

Potential of the trifunctional bispecific antibody surek depends on dendritic cells: rationale for a new approach of tumor immunotherapy.  

PubMed

Trifunctional bispecific antibodies (trAbs) used in tumor immunotherapy have the unique ability to recruit T cells toward antigens on the tumor cell surface and, moreover, to activate accessory cells through their immunoglobulin Fc region interacting with activating Fc? receptors. This scenario gives rise to additional costimulatory signals required for T cell-mediated tumor cell destruction and induction of an immunologic memory. Here we show in an in vitro system that most effective trAb-dependent T-cell activation and tumor cell elimination are achieved in the presence of dendritic cells (DCs). On the basis of these findings, we devise a novel approach of cancer immunotherapy that combines the specific advantages of trAbs with those of DC-based vaccination. Simultaneous delivery of trAbs and in vitro differentiated DCs resulted in a markedly improved tumor rejection in a murine melanoma model compared with monotherapy. PMID:23552725

Eissler, Nina; Mysliwietz, Josef; Deppisch, Nina; Ruf, Peter; Lindhofer, Horst; Mocikat, Ralph

2013-01-01

195

Activation of human complement by totally human monoclonal antibodies  

Microsoft Academic Search

A uniquely developed series of totally§§The term “totally” is used to distinguish these antibodies from humanized immunoglobulins or monoclonals produced from mouse\\/human heterohybridomas. human monoclonal antibodies (mAbs) were examined for their complement fixing properties in comparison to human myeloma preparations and to commercially available human polyclonal immunoglobulins. C3b and C4b deposition was measured using a kinetic ELISA technique. When the

Susanne L. Dillman; Anthony J. Strelkauskas; Helene R. Su; Robert J. Boackle

1995-01-01

196

Use of commercially available monoclonal antibodies for immunoenzyme double staining  

Microsoft Academic Search

Summary  An immunoenzyme double-staining method for the simultaneous detection of two cellular epitopes, using commercially available mouse monoclonal antibodies, is described. The method employs a combination of the suppression of endogenous biotin and two successive indirect techniques with a blocking step in between. The first indirect method involves an unlabelled monoclonal antibody followed by an alkaline phosphatase-conjugated goat anti-mouse immunoglobulin. After

C. M. Van Der Loos; J. J. Van Den Oord; P. K. Das; H. J. Houthoff

1988-01-01

197

Cutaneous adverse reactions to therapeutic monoclonal antibodies for cancer  

Microsoft Academic Search

Advances in molecular biology have led to the successful development of targeted monoclonal antibodies to several types of\\u000a malignancies. By June 2007, nine therapeutic monoclonal antibodies had been approved by the US Food and Drug Administration\\u000a to treat various human cancers. In general, the adverse reactions of these agents have been milder than their cytotoxic chemotherapy\\u000a counterparts, but side effects

Patricia L. Myskowski; Allan C. Halpern

2008-01-01

198

Cutaneous adverse reactions to therapeutic monoclonal antibodies for cancer  

Microsoft Academic Search

Advances in molecular biology have led to the successful development of targeted monoclonal antibodies to several types of\\u000a malignancies. By June 2007, nine therapeutic monoclonal antibodies had been approved by the US Food and Drug Administration\\u000a to treat various human cancers. In general, the adverse reactions of these agents have been milder than their cytotoxic chemotherapy\\u000a counterparts, but side effects

Patricia L. Myskowski; Allan C. Halpern

2009-01-01

199

Monoclonal Antibodies for Systemic Lupus Erythematosus (SLE) †  

PubMed Central

A number of monoclonal antibodies (mAb) are now under investigation in clinical trials to assess their potential role in Systemic Lupus Erythematosus (SLE). The most frequently used mAb is rituximab, which is directed against CD20, a membrane protein expressed on B lymphocytes. Uncontrolled trials reported an improvement of SLE activity in non-renal patients and other studies even reported an improvement of severe lupus nephritis unresponsive to conventional treatments. However two randomized trials failed to show the superiority of rituximab over conventional treatment in non renal SLE and in lupus nephritis. Preliminary trials reported promising results with epratuzumab, a humanized mAb directed against CD22, and with belimumab, a human mAb that specifically recognizes and inhibits the biological activity of BLyS a cytokine of the tumor-necrosis-factor (TNF) ligand superfamily. Other clinical trials with mAb directed against TNF-alpha, interleukin-10 (Il-10), Il-6, CD154, CD40 ligand, IL-18 or complement component C5 are under way. At present, however, in spite of good results reported by some studies, no firm conclusion on the risk-benefit profile of these mAbs in patients with SLE can be drawn from the available studies.

Ponticelli, Claudio; Moroni, Gabriella

2010-01-01

200

Radiolabeled monoclonal antibodies for medical applications  

SciTech Connect

Hybridoma technology can be used to develop monoclonal antibodies (MAbs) that react with specific structures present on the surface of a variety of cell populations. Radioiodinated MAbs are now used routinely in in vitro tests for the diagnosis of ovarian, colorectal, and other cancers. The availability of this methodology has also had a major impact on the current directions of research in nuclear medicine because of the potential for utilizing MAbs for the selective delivery of nuclides to tumors or other biologic targets. When labeled with an appropriate gamma emitter, MAbs can be used to define the location and extent of cancer and other diseases through the use of conventional planar imaging, single photon emission computed tomography (SPET) or positron emission tomography (PET). An exciting possibility is to use MAb imaging as a prelude to the therapeutic application of MAbs labeled with nuclides decaying by the emission of either beta or alpha particles. Radioimmunotherapy shows promise as an approach for delivering curative doses of radiation to malignant cell populations without irreversibly damaging neighboring normal tissues. Numerous tumor-imaging studies using MAbs labeled with {sup 131}I, {sup 111}In, {sup 123}I, and {sup 99m}Tc have been performed in patients with a variety of malignancies, including melanoma, lymphoma, and ovarian, breast, and colorectal carcinomas.

Zalutsky, M.R.; Garg, P.K.; Vaidyanathan, G. (Duke Univ. Medical Center, Durham, NC (United States))

1992-01-01

201

Monoclonal antibodies against plant cell wall polysaccharides  

SciTech Connect

Monoclonal antibodies (McAbs) are useful tools to probe the structure of plant cell wall polysaccharides and to localize these polysaccharides in plant cells and tissues. Murine McAbs were generated against the pectic polysaccharide, rhamnogalacturonan I (RG-I), isolated from suspension-cultured sycamore cells. The McAbs that were obtained were grouped into three classes based upon their reactivities with a variety of plant polysaccharides and membrane glycoproteins. Eleven McAbs (Class I) recognize epitope(s) that appear to be immunodominant and are found in RG-I from sycamore and maize, citrus pectin, polygalacturonic acid, and membrane glycoproteins from suspension-cultured cells of sycamore, maize, tobacco, parsley, and soybean. A second group of five McAbs (Class II) recognize epitope(s) present in sycamore RG-I, but do not bind to any of the other polysaccharides or glycoproteins recognized by Class I. Lastly, one McAb (Class III) reacts with sycamore RG-I, sycamore and tamarind xyloglucan, and sycamore and rice glucuronoarabinoxylan, but does not bind to maize RG-I, polygalacturonic acid or the plant membrane glycoproteins recognized by Class I. McAbs in Classes II and III are likely to be useful in studies of the structure, biosynthesis and localization of plant cell wall polysaccharides.

Hahn, M.G.; Bucheli, E.; Darvill, A.; Albersheim, P. (Univ. of Georgia, Athens (USA))

1989-04-01

202

Monoclonal antibodies as therapeutics in human malignancies.  

PubMed

Monoclonal antibodies (mAbs) are a proven effective therapeutic modality in human malignancy. Several mAbs are approved to targets critical in aberrant oncogenic signaling within tumors and their microenvironment. These targets include secreted ligands (e.g., VEGF and HGH), their receptors (e.g., HER2 and VEGFR2), cell surface counter receptors and their receptor-bound ligands (e.g., PD1 and PD1L, respectively). The ability to genetically engineer the structure and/or functions of mAbs has significantly improved their effectiveness. Furthermore, advances in gene expression profiling, proteomics, deep sequencing and deciphering of complex signaling networks have revealed novel therapeutic targets. We review target selection, approved indications and the rationale for mAb utilization in solid and hematologic malignancies. We also discuss novel mAbs in early- and late-phase clinical trials that are likely to change the natural history of disease and improve survival. The future challenge is to design mAb-based novel trial designs for diagnostics and therapeutics for human malignancies. PMID:24754592

Pandey, Manjari; Mahadevan, Daruka

2014-03-01

203

Perspectives on anti-HER monoclonal antibodies.  

PubMed

The ability of Herceptin to prolong survival in women with HER2-overexpressing breast tumors has proven the concept of using humanized or chimeric monoclonal antibodies (MAbs) for cancer therapy. MAbs have been developed that are specific for many tumorigenic molecules and receptors. They can potentially be used to treat a range of solid tumors. Among the most promising targets for therapy are members of the human epidermal growth factor receptor (HER/ErbB) family, particularly HER1 and HER2. Several MAbs have been produced that are directed against HER1. One of these agents, cetuximab (Erbitux), is now advanced in clinical development. HER2 is also a key target and methods are being investigated to maximize the effect of using MAbs to inhibit this receptor. One approach aims to augment the efficacy of trastuzumab (Herceptin) by coupling it to a chemotherapeutic agent, thus enabling the delivery of cytotoxic therapy at a cellular level. Another opportunity is based on research that shows that HER2 acts as a dimerization partner for other HER receptors and consequently is important in HER-ligand-dependent tumor growth. Therefore, anti-HER2 MAbs that inhibit the association of HER2 with other HER family members have the potential to be highly effective. This article reviews some of these alternative approaches to MAb-based anti-HER therapy that will hopefully improve treatment outcome for patients with a range of solid tumors. PMID:12422051

Ranson, Malcolm; Sliwkowski, Mark X

2002-01-01

204

Monoclonal antibodies: new agents for cancer detection and targeted therapy  

SciTech Connect

Antibodies directed against markers on cancer cells are gaining in importance for the purpose of targeting diagnostic and therapeutic agents. In the past, this approach has had very limited success principally because the classical methods for producing antibodies from blood serum of animals immunized with cancer cells or extracts were unsatisfactory. The situation has changed dramatically since 1975 following the design of procedures for 'immortalizing' antibody-producing cells (lymphocytes) by fusing them with cultured myeloma cells to form hybridomas which continuously secrete antibodies. Since these hybridomas produce antibodies coded for by a single antibody-producing cell, the antibodies are called monoclonal. Building on these advances in biomedical research, it is now possible to reproducibly manufacture monoclonal antibodies on a scale suitable for use in cancer detection and therapy.

Baldwin, R.W.; Byers, V.S. (Univ. of Nottingham (United Kingdom))

1991-01-01

205

Boronated monoclonal antibody conjugates for neutron capture therapy  

SciTech Connect

This paper describes the effectiveness of /sup 10/B-labeled monoclonal antibodies against Colo-38 human melanoma in vitro. The authors obtained high boron to antibody ratios while maintaining antibody activity by using dextran intermediate carriers to link /sup 10/B to the antibody. They developed a double cell quasi-competitive binding bioassay to minimize the effects of nonspecific binding of boronated complexes to cells. 1 fig., 2 tabs.

Borg, D.C.; Elmore, J.J. Jr.; Ferrone, S.

1986-01-01

206

Characterization and utilization of a monoclonal antibody against pancreatic carcinoma  

SciTech Connect

A monoclonal antibody was produced against a human pancreatic adenocarcinoma line and was found to react with several different human carcinomas by immunoperoxidase staining of fixed tissues. The original cells used to generate the monoclonal antibody were treated with detergent to lyse the cell membrane. A membrane associated protein of molecular weight 35kD was isolated from this detergent lysed preparation and found to be recognized by the monoclonal antibody. The binding constant of the antigen antibody reaction on the cells is 5 x 10{sup {minus}5}. It was further determined that there are 700,000 binding sites per cell. Kinetics of the antigen-antibody reaction under several conditions were also explored.

Kurtzman, S.H.; Sindelar, W.F.; Atcher, R.W.; Mitchell, J.B.; DeGraff, W.G.; Gamson, J.; Russo, A. [National Cancer Institute, Bethesda, MD (United States); Friedman, A.M.; Hines, J.J. [Argonne National Lab., IL (United States)

1994-10-01

207

Monoclonal antibody therapy in multiple sclerosis  

PubMed Central

Therapeutic approaches to multiple sclerosis (MS) are based on altering the functions of the immune system, either by using broad immunosuppressive drugs used for transplantation rejection and rheumatology, or by modulating them more discreetly with beta interferon and synthetic amino-acid copolymers. These strategies are only partially successful, have important safety and tolerability limitations, and have shown to be mostly effective in earlier stages of the disease, in which acute relapses dominate the clinical picture. For progressive phenotypes of MS there are currently no effective therapeutic options. As very specific and potent immunosuppressive agents, monoclonal antibodies (mAbs) may offer considerable advantages over other therapies for MS. During the last decade, anti-a4 integrin natalizumab became the first approved mAb for treatment of relapsing MS, after convincingly demonstrating clinically significant effects on two large Phase 3 trials. Moreover, the concept of disease remission was introduced for the first time to describe patients who show no signs of clinical or imaging markers of disease activity during therapy with natalizumab. Of the mAbs under development for MS, alemtuzumab and rituximab have also shown promising evidence of effectiveness and potentially expanded the therapeutic horizon to reversal of disease progression in early relapsing patients and progressive patients who previously had not been studied. However, the appearance of progressive multifocal leukoencephalopathy (PML) in natalizumab-treated MS patients, as well as in patients with lymphoma, lupus and rheumatoid arthritis, treated with rituximab and autoimmune-type complications in alemtuzumab-treated MS patients underlines the fact that extended efficacy comes with significant clinical risks. The challenge is then how best to utilize therapies that have evidently superior efficacy in a chronic disease of young adults to obtain the best benefit-risk ratio and how to monitor and prevent emergent safety concerns.

2010-01-01

208

Function-blocking antithrombospondin-1 monoclonal antibodies  

PubMed Central

Summary Background Thrombospondin-1 (TSP-1) has been implicated in many different processes based in part on inhibitory activities of anti-TSP-1 monoclonal antibodies (mAbs). Objective To map epitopes of 13 anti-TSP-1 mAbs to individual modules or groups of modules spanning TSP-1 and the closely related TSP-2 homolog. Results The mapping has led to assignment or reassignment of the epitopes of four mAbs, refinement of the epitopes of six mAbs, and confirmation of the epitopes of the remaining three mAbs. ESTs10, P12, and MA-II map to the N-terminal domain; 5G11, TSP127.6, and ESTs12 to the third properdin module; C6.7, HB8432, and P10 to epidermal growth factor (EGF)-like modules 1 and/or 2; and A6.1, mAb133, MA-I, and D4.6 to the calcium-binding wire module. A6.1, which recognizes a region of the wire that is identical in mouse and human TSP-1, reacts with TSP-1 from both species, and also reacts weakly with human TSP-2. Two other mouse antihuman TSP-1 mAbs, A4.1 and D4.6, also react with mouse TSP-1. Conclusions Consideration of previous literature and mapping of epitopes of inhibitory mAbs suggest that biological activities are present throughout TSP-1, including the EGF-like modules that have not been implicated in the past. Because the epitopes for 10 of the antibodies likely are within 18 nm of one another in calcium-replete TSP-1, some of the inhibitory effects may result from steric hindrance. Such seems to be the case for mAb133, which binds the calcium-binding wire but is still able to interfere with the activation of latent TGF-? by the properdin modules.

ANNIS, D. S.; MURPHY-ULLRICH, J. E.; MOSHER, D. F.

2006-01-01

209

Monoclonal antibodies: versatile platforms for cancer immunotherapy  

Microsoft Academic Search

Antibodies are important therapeutic agents for cancer. Recently, it has become clear that antibodies possess several clinically relevant mechanisms of action. Many clinically useful antibodies can manipulate tumour-related signalling. In addition, antibodies exhibit various immunomodulatory properties and, by directly activating or inhibiting molecules of the immune system, antibodies can promote the induction of antitumour immune responses. These immunomodulatory properties can

Rishi Surana; Shangzi Wang; Louis M. Weiner

2010-01-01

210

Evidence of a saturable hepatic receptor for mouse monoclonal antibodies  

Microsoft Academic Search

Monoclonal antibodies (MAb) can be labeled with I-123 at high specific activities, so that large amounts of radioactivity attached to small amounts of protein can be injected for radioimmunoimaging. This conserves antibody and decreases the opportunity for foreign protein reactions and target tissue binding site saturation. In order to assess the effects on pharmacokinetics and imaging, the authors administered microgram

G. L. De Nardo; S. J. De Nardo; J. S. Peng; L. F. OGrady; S. L. Mills; A. L. Epstein; R. D. Cardiff

1985-01-01

211

Research and Diagnostic Applications of Monoclonal Antibodies to 'Coccidioides immitis'.  

National Technical Information Service (NTIS)

The authors have been assembling a panel of mouse monoclonal antibodies to antigens on the surface of endospores and spherules, and in culture filtrates and soluble extracts of spherule-phase C. immitis. These antibodies are being used to develop antigen-...

A. E. Karu D. J. Gennevois J. W. Hoffman S. J. Kraeger H. B. Levine

1985-01-01

212

Palladium-109 labeled anti-melanoma monoclonal antibodies  

DOEpatents

The invention consists of new monoclonal antibodies labelled with Palladium 109, a beta-emitting radionuclide, the method of preparing this material, and its use in the radiotherapy of melanoma. The antibodies are chelate-conjugated and demonstrate a high uptake in melanomas. (ACR)

Srivastava, S.C.; Fawwaz, R.A.; Ferrone, S.

1984-04-30

213

Human Monoclonal Antibodies for Neutralization of Botulinum Neurotoxin.  

National Technical Information Service (NTIS)

The purpose of this work is to generate neutralizing human monoclonal antibodies to Botulinum neurotoxins (BoNT) A, B, and E. To generate a large panel of antibodies, mice transgenic for the human immunoglobulin were immunized with BoNT/A, B, and E bindin...

J. D. Marks

2000-01-01

214

Neutralizing monoclonal antibodies directed to infectious bovine rhinotracheitis virus.  

PubMed

Infectious bovine rhinotracheitis virus (IBRV) has been shown in this report to have thirty-three polypeptides. Ten of the eleven polypeptides which can be labeled with (3H)-glucosamine are located on the surface of the virus since they can be surface labeled with sodium boro(3H)hydride. In order to define the immunologically important viral proteins, monoclonal antibodies were prepared against the virus and selected for their ability to neutralize infectivity. Four such hybridoma lines were obtained for characterization of the antigens that elicit neutralizing antibodies. The viral polypeptides were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of each monoclonal antibody was determined by "Western" blot analysis and/or by immunoprecipitation of (35S)-methionine and (3H)-glucosamine labeled infected cell lysates by the monoclonal antibodies. One monoclonal antibody reacted with two glycoproteins, gp135 and gp78a, on the "Western" blot but immunoprecipitated three glycoproteins, gp135, gp78a, and gp54 from labeled infected cell lysates. The other three monoclonal antibodies immunoprecipitated a single glycoprotein, gp78b, from (3H)-glucosamine labeled infected cell lysates but not from (35S)-methionine labeled infected cell lysates. PMID:3010905

Chang, L W; Zee, Y C; Pritchett, R F; Ardans, A A

1986-01-01

215

Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies.  

PubMed

Antigen specific llama VHH antibody fragments were compared to antigen specific mouse monoclonal antibodies with respect to specificity, affinity and stability. The llama VHH antibody fragments and the mouse monoclonal antibodies investigated were shown to be highly specific for the protein antigen hCG or the hapten antigen RR-6. The affinity of the interaction between monovalent llama VHH antibody fragments and their antigen is close to the nanomolar range, similar to the bivalent mouse monoclonal antibodies studied. Llama VHH antibody fragments are similar to mouse monoclonal antibodies with respect to antigen binding in the presence of ammonium thiocyanate and ethanol. The results show that relative to antigen specific mouse monoclonal antibodies, antigen specific llama VHH fragments are extremely temperature stable. Two out of six llama VHHs are able to bind antigen specifically at temperatures as high as 90 degrees C, whereas four out of four mouse monoclonal antibodies are not functional at this temperature. Together with the finding that llama VHH fragments can be produced at high yield in Saccharomyces cerevisiae, these findings indicate that in the near future antigen specific llama VHH fragments can be used in for antibodies unexpected products and processes. PMID:10209277

van der Linden, R H; Frenken, L G; de Geus, B; Harmsen, M M; Ruuls, R C; Stok, W; de Ron, L; Wilson, S; Davis, P; Verrips, C T

1999-04-12

216

A bispecific antibody to factors IXa and X restores factor VIII hemostatic activity in a hemophilia A model.  

PubMed

Hemophilia A is a bleeding disorder resulting from coagulation factor VIII (FVIII) deficiency. Exogenously provided FVIII effectively reduces bleeding complications in patients with severe hemophilia A. In approximately 30% of such patients, however, the 'foreignness' of the FVIII molecule causes them to develop inhibitory antibodies against FVIII (inhibitors), precluding FVIII treatment in this set of patients. Moreover, the poor pharmacokinetics of FVIII, attributed to low subcutaneous bioavailability and a short half-life of 0.5 d, necessitates frequent intravenous injections. To overcome these drawbacks, we generated a humanized bispecific antibody to factor IXa (FIXa) and factor X (FX), termed hBS23, that places these two factors into spatially appropriate positions and mimics the cofactor function of FVIII. hBS23 exerted coagulation activity in FVIII-deficient plasma, even in the presence of inhibitors, and showed in vivo hemostatic activity in a nonhuman primate model of acquired hemophilia A. Notably, hBS23 had high subcutaneous bioavailability and a 2-week half-life and would not be expected to elicit the development of FVIII-specific inhibitory antibodies, as its molecular structure, and hence antigenicity, differs from that of FVIII. A long-acting, subcutaneously injectable agent that is unaffected by the presence of inhibitors could markedly reduce the burden of care for the treatment of hemophilia A. PMID:23023498

Kitazawa, Takehisa; Igawa, Tomoyuki; Sampei, Zenjiro; Muto, Atsushi; Kojima, Tetsuo; Soeda, Tetsuhiro; Yoshihashi, Kazutaka; Okuyama-Nishida, Yukiko; Saito, Hiroyuki; Tsunoda, Hiroyuki; Suzuki, Tsukasa; Adachi, Hideki; Miyazaki, Taro; Ishii, Shinya; Kamata-Sakurai, Mika; Iida, Takeo; Harada, Aya; Esaki, Keiko; Funaki, Miho; Moriyama, Chifumi; Tanaka, Eriko; Kikuchi, Yasufumi; Wakabayashi, Tetsuya; Wada, Manabu; Goto, Masaaki; Toyoda, Takeshi; Ueyama, Atsunori; Suzuki, Sachiyo; Haraya, Kenta; Tachibana, Tatsuhiko; Kawabe, Yoshiki; Shima, Midori; Yoshioka, Akira; Hattori, Kunihiro

2012-10-01

217

Purification of hemagglutinin from Haemophilus paragallinarum using monoclonal antibody.  

PubMed

The purification of hemagglutinin from Haemophilus paragallinarum was attempted using affinity chromatography with monoclonal antibody. The antigen eluted from the affinity column using potassium thiocyanate buffer agglutinated chicken erythrocytes. In immunoblotting of the eluted antigen, a single band with monoclonal antibody was found as well as the crude antigen. When the chickens were immunized with the eluted antigen, they produced the hemagglutination inhibition (HI) antibody, and they showed protection against challenged exposure with H. paragallinarum strain 221. These results indicated that the HA antigen of H. paragallinarum was a protective antigen. PMID:8451834

Takagi, M; Hirayama, N; Simazaki, T; Taguchi, K; Yamaoka, R; Ohta, S

1993-02-01

218

A perspective of monoclonal antibodies: Past, present, and future  

SciTech Connect

In 1975, the development of the technique to produce monoclonal antibodies revolutionized the approach to cancer detection and therapy. Hundreds of monoclonal antibodies to the epitopes of tumor cells have been produced, providing more specific tools for probing the cellular elements of cancer. At the same time, these tools have disclosed greater complexity in the character of these cells and stimulated further investigation. Although there are antibodies to specific epitopes of neoplastic cells, this purity has not provided the improved detection and therapy of cancer first expected. Technical manipulations have provided limited improvement in results, but more sophisticated techniques, such as biologic response modifiers, may be required to attain clinical results that can be universally applied. The intense research in monoclonal antibodies and their application does offer promise that the goal of improved cancer detection and therapy will be forthcoming. 58 references.

DeLand, F.H. (Veterans Administration Medical Center, Syracuse, NY (USA))

1989-07-01

219

Highly Efficient Elimination of Colorectal Tumor-Initiating Cells by an EpCAM\\/CD3-Bispecific Antibody Engaging Human T Cells  

Microsoft Academic Search

With their resistance to genotoxic and anti-proliferative drugs and potential to grow tumors and metastases from very few cells, cancer stem or tumor-initiating cells (TICs) are a severe limitation for the treatment of cancer by conventional therapies. Here, we explored whether human T cells that are redirected via an EpCAM\\/CD3-bispecific antibody called MT110 can lyse colorectal TICs and prevent tumor

Ines Herrmann; Patrick A. Baeuerle; Matthias Friedrich; Alexander Murr; Susanne Filusch; Dominik Rüttinger; Mariam W. Majdoub; Sherven Sharma; Peter Kufer; Tobias Raum; Markus Münz; Dominik Hartl

2010-01-01

220

The CEA\\/CD3-Bispecific Antibody MEDI565 (MT111) Binds a Nonlinear Epitope in the Full-Length but Not a Short Splice Variant of CEA  

Microsoft Academic Search

MEDI-565 (also known as MT111) is a bispecific T-cell engager (BiTE®) antibody in development for the treatment of patients with cancers expressing carcinoembryonic antigen (CEA). MEDI-565 binds CEA on cancer cells and CD3 on T cells to induce T-cell mediated killing of cancer cells. To understand the molecular basis of human CEA recognition by MEDI-565 and how polymorphisms and spliced

Li Peng; Michael D. Oberst; Jiaqi Huang; Philip Brohawn; Chris Morehouse; Kristen Lekstrom; Patrick A. Baeuerle; Herren Wu; Yihong Yao; Steven R. Coats; William Dall’Acqua; Melissa Damschroder; Scott A. Hammond

2012-01-01

221

Preclinical Evaluation of Multistep Targeting of Diasialoganglioside GD2 Using an IgG-scFv Bispecific Antibody with High Affinity for GD2 and DOTA Metal Complex.  

PubMed

Bispecific antibodies (BsAb) have proven to be useful targeting vectors for pretargeted radioimmunotherapy (PRIT). We sought to overcome key PRIT limitations such as high renal radiation exposure and immunogenicity (e.g., of streptavidin-antibody fusions), to advance clinical translation of this PRIT strategy for diasialoganglioside GD2-positive [GD2(+)] tumors. For this purpose, an IgG-scFv BsAb was engineered using the sequences for the anti-GD2 humanized monoclonal antibody hu3F8 and C825, a murine scFv antibody with high affinity for the chelator 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) complexed with ?-particle-emitting radiometals such as (177)Lu and (90)Y. A three-step regimen, including hu3F8-C825, a dextran-based clearing agent, and p-aminobenzyl-DOTA radiolabeled with (177)Lu (as (177)Lu-DOTA-Bn; t1/2 = 6.71 days), was optimized in immunocompromised mice carrying subcutaneous human GD2(+) neuroblastoma (NB) xenografts. Absorbed doses for tumor and normal tissues were approximately 85 cGy/MBq and ?3.7 cGy/MBq, respectively, with therapeutic indices (TI) of 142 for blood and 23 for kidney. A therapy study (n = 5/group; tumor volume, 240 ± 160 mm(3)) with three successive PRIT cycles (total (177)Lu: ?33 MBq; tumor dose ?3,400 cGy), revealed complete tumor response in 5 of 5 animals, with no recurrence up to 28 days after treatment. Tumor ablation was confirmed histologically in 4 of 5 mice, and normal organs showed minimal overall toxicities. All nontreated mice required sacrifice within 12 days (>1.0-cm(3) tumor volume). We conclude that this novel anti-GD2 PRIT approach has sufficient TI to successfully ablate subcutaneous GD2(+)-NB in mice while sparing kidney and bone marrow. Mol Cancer Ther; 13(7); 1803-12. ©2014 AACR. PMID:24944121

Cheal, Sarah M; Xu, Hong; Guo, Hong-Fen; Zanzonico, Pat B; Larson, Steven M; Cheung, Nai-Kong

2014-07-01

222

Monoclonal antibody for calcitriol (1 alpha,25-dihydroxyvitamin D3).  

PubMed

Hybridoma cell lines secreting antibodies for vitamin D3 metabolites have been generated by fusing splenocytes from BALB/c mice immunized with 3 beta-glutaryl-25-hydroxyvitamin D3 conjugated to bovine serum albumin (3 beta-glu-25-OH-D3-BSA) and Sp2/O-Ag14 myeloma cells. Purification of monoclonal antibodies from culture media or ascites fluids was accomplished by procedures including affinity chromatography on Protein A-Sepharose 4B. Each monoclonal antibody was analyzed as to its affinity and specificity by equilibrium dialysis and an enzyme immunoassay (EIA) based on a double antibody system. It was demonstrated that clone 1C2-60 produced an antibody highly specific to 1 alpha,25-dihydroxyvitamin D3 (calcitriol), and the clone 2B3-66 antibody was reactive to 25-hydroxyvitamin D3 and similar structural compounds. These two monoclonal antibodies produced by 1C2-60 and 2B3-66 were determined to belong to the IgG2a class, and their affinity constants (Ka) with 3 beta-glu-25-OH-D3 were demonstrated to be 3.6 X 10(9) M-1 and 2.9 X 10(9) M-1, respectively, at 4 degrees C. The characteristics of these monoclonal antibodies were compared with those of conventional antibodies raised in mice and rabbits. Finally, by using monoclonal antibody 1C2-60, a sensitive EIA has been developed that can detect 10 pg of calcitriol. PMID:3908452

Yamamoto, I; Matsuura, E

1985-10-01

223

Monoclonal Antibodies in Gynecological Cancer: A Critical Point of View  

PubMed Central

During the last decades, several improvements in treating gynecological malignancies have been achieved. In particular, target therapies, mostly monoclonal antibodies, have emerged as an attractive option for the treatment of these malignancies. In fact, various molecular-targeted agents have been developed for a variety of malignancies with the objective to interfere with a precise tumor associated receptor, essential for cancer cell survival or proliferation, blocking its function, of the cancer cells. Alternatively, monoclonal antibodies have been developed to block immune suppression or enhance functions of immune effector cells. So far, several monoclonal antibodies have been tested for clinical efficacy for the treatment of gynecological cancers. Antibodies against Vascular Endothelial Growth Factor (VEGF) and Epidermal Growth Factor Receptor (EGFR) have been used in different neoplasms such as ovarian and cervical cancer. Catumazumab, a bivalent antibody against CD3 and EpCAM, is effective in the treatment of neoplastic ascites. Other antibodies are peculiar for specific cancer-associated antigen such as Oregovomab against CA125 or Farletuzumab against the folate receptor. Here we describe the preclinical and clinical experience gained up to now with monoclonal antibodies in tumors of the female genital tract and trace future therapeutic and research venues.

Bellati, Filippo; Napoletano, Chiara; Gasparri, Maria Luisa; Visconti, Valeria; Zizzari, Ilaria Grazia; Ruscito, Ilary; Caccetta, Jlenia; Rughetti, Aurelia; Benedetti-Panici, Pierluigi; Nuti, Marianna

2011-01-01

224

Breast cancer immunotherapy: monoclonal antibodies and peptide-based vaccines.  

PubMed

Recently, immunotherapy has emerged as a treatment strategy in the adjuvant setting of breast cancer. In this review, monoclonal antibodies in passive and peptide-based vaccines, as one of the most commonly studied in active immunotherapy approaches, are discussed. Trastuzumab, a monoclonal antibody against HER-2/neu, has demonstrated considerable efficacy. However, resistance to trastuzumab has led to development of many targeted therapies which have been examined in clinical trials. Monoclonal antibodies against immune-checkpoint molecules that are dysregulated by tumors as an immune resistance mechanism are also explained in this review. Additionally, monoclonal antibodies with the ability to target breast cancer stem cells that play a role in cancer recurrence are mentioned. Here, clinical trials of HER-2/neu B and T cells, MUC1 and hTERT cancer peptide vaccines are also presented. In addition, various strategies for enhancing vaccine efficacy including combination with monoclonal antibodies and using different delivery systems for peptide/protein-based vaccine are described. PMID:24867051

Mohit, Elham; Hashemi, Atieh; Allahyari, Mojgan

2014-07-01

225

Chemotherapy Combinations With Monoclonal Antibodies in Non-Hodgkin's Lymphoma  

PubMed Central

Although the use of monoclonal antibodies as single agents has had a tremendous impact on the care of patients with non-Hodgkin’s lymphoma (NHL), the greatest benefit has been generated by the addition of monoclonal antibodies to conventional cytotoxic chemotherapy. Rituximab is the monoclonal antibody responsible for all clinical improvement noted to date. The addition of rituximab to cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy (R-CHOP regimen) improves the response rate, progression-free survival (PFS), and overall survival (OS) in diffuse large B-cell lymphoma (DLBCL). Adding rituximab to CHOP chemotherapy improves response rates and PFS in mantle cell lymphoma (MCL). Finally, the addition of rituximab to a variety of chemotherapy regimens improves the response rates, PFS, and OS in follicular lymphoma (FL). Several other (epratuzumab, bevacizumab, alemtuzumab) monoclonal antibody–chemotherapy combinations are currently under study in NHL. This review will summarize the data supporting the addition of rituximab to chemotherapy in NHL and discuss preliminary data regarding the use of other monoclonal antibodies in combination with chemotherapy.

Kahl, Brad

2010-01-01

226

Eradication of Tumors from a Human Colon Cancer Cell Line and from Ovarian Cancer Metastases in Immunodeficient Mice by a Single-Chain Ep-CAM-\\/CD3-Bispecific Antibody Construct  

Microsoft Academic Search

Bispecific T-cell engager (BiTE) are a class of bispecific single- chain antibodies that can very effectively redirect cytotoxic T cells for killing of tumor target cells. Here, we have assessed the in vivo efficacy of one representative, called bscEp- CAMxCD3, with specificity for tumors overexpressing epithe- lial cell adhesion molecule (Ep-CAM) in human xenograft models. Cells of the human colon

Bernd Schlereth; Iduna Fichtner; Grit Lorenczewski; Petra Kleindienst; Klaus Brischwein; Antonio da Silva; Peter Kufer; Ralf Lutterbuese; Ilse Junghahn; Sabine Kasimir-Bauer; Pauline Wimberger; Rainer Kimmig; Patrick A. Baeuerle

227

Production of type II collagen specific monoclonal antibodies.  

PubMed

Immunoassays are used for the specific measurement of type II collagen, a major cartilage protein, which is lost in osteoarthritic joints. Poor immunogenicity and species dependent immune response to type II collagen make it difficult to obtain specific antibodies required for immunoassay development. In addition, type II collagen antibodies exhibit reactivity to structurally dissimilar antigens such as actin, myoglobin, thyroglobulin and ssDNA, complicating the isolation of specific antibodies. It is therefore necessary to characterize the antibody reactivity against both noncollagenous antigens and different collagen types. In this study, immune response to type II collagen was improved by conjugation to carrier proteins, KLH and BSA. Hybridomas were generated by fusions of lymphocytes derived from lymph nodes or spleens with X63-653-Ag8 myeloma cells. Compared to spleens, the utilization of lymph nodes as a source of lymphocytes resulted in a 23% higher number of hybridomas secreting type II collagen antibodies. Hybridomas secreting polyreactive antibodies were identified based on their reactivity to thyroglobulin and eliminated. Extensive testing of the remaining monoclonal antibodies with other structurally dissimilar antigens and various types of collagen for reactivity, allowed us to isolate specific monoclonal antibodies to type II collagen. We emphasize the importance of characterization of the reactivity of type II collagen monoclonal antibodies before employing them for immunoassays. PMID:8194857

Srinivas, G R; Chichester, C O; Barrach, H J; Pillai, V; Matoney, A L

1994-03-01

228

Population pharmacokinetics of therapeutic monoclonal antibodies.  

PubMed

A growing number of population pharmacokinetic analyses of therapeutic monoclonal antibodies (mAbs) have been published in the scientific literature. The aims of this article are to summarize the findings from these studies and to relate the findings to the general pharmacokinetic and structural characteristics of therapeutic mAbs. A two-compartment model was used in the majority of the population analyses to describe the disposition of the mAb. Population estimates of the volumes of distribution in the central (V(1)) and peripheral (V(2)) compartments were typically small, with median (range) values of 3.1 (2.4-5.5) L and 2.8 (1.3-6.8) L, respectively. The estimated between-subject variability in the V(1) was usually moderate, with a median (range) coefficient of variation (CV) of 26% (12-84%). Between-subject variability in other distribution-related parameters such as the V(2) and intercompartmental clearance were often not estimated. Although the pharmacokinetic models used most frequently in the population analyses were models with linear clearance, other models with nonlinear, or parallel linear and nonlinear clearance pathways were also applied, as many therapeutic mAbs are eliminated via saturable target-mediated mechanisms. Population estimates of the maximum elimination rate (V(max)) and the mAb concentration at which elimination was at half maximum for Michaelis-Menten-type elimination pathways varied considerably among the different therapeutic mAbs. However, estimates of the total clearance (CL) of mAbs with linear clearance characteristics and of the clearance of mAbs via the linear clearance pathway (CL(L)) with parallel linear and nonlinear clearance were quite similar for the different mAbs and typically ranged from 0.2 to 0.5 L/day, which is relatively close to the estimated clearance of endogenous IgG of 0.21 L/day. The between-subject variability in the V(max), CL and CL(L) was moderate to high, with estimated CVs ranging from 15% to 65%. Measures of body size were the covariates most commonly identified as influencing the pharmacokinetics of therapeutic mAbs. In summary, many features of the population pharmacokinetics of currently used therapeutic mAbs are similar, despite differences in their pharmacological targets and studied patient populations. PMID:20818831

Dirks, Nathanael L; Meibohm, Bernd

2010-10-01

229

IgG2a Monoclonal Antibodies Inhibit Human Tumor Growth through Interaction with Effector Cells  

NASA Astrophysics Data System (ADS)

Monoclonal antibodies of IgG2a isotype specifically inhibited growth of human tumors in nude mice. Twentythree monoclonal antibodies of other isotypes showed no tumoricidal reactivity. Complement depletion of nude mice had no effect on tumor suppression by monoclonal antibody. The role of T and killer cells as mediators of the monoclonal antibody effect in nude mice was virtually excluded. On the other hand, macrophages were strongly incriminated as effector cells because silica treatment of nude mice abolished the tumoricidal effect of monoclonal antibody. IgG2a monoclonal antibody-dependent macrophagemediated cytotoxicity assays with human tumor cells in culture resulted in specific lysis of tumor cells.

Herlyn, Dorothee; Koprowski, Hilary

1982-08-01

230

Potent inhibition of local and disseminated tumor growth in immunocompetent mouse models by a bispecific antibody construct specific for Murine CD3.  

PubMed

Bispecific single-chain antibody constructs specific for human CD3 have been extensively studied for antitumor activity in human xenograft models using severe combined immunodeficient mice supplemented with human T cells. High efficacy at low effector-to-target ratios, independence of T cell costimuli and a potent activation of previously unstimulated polyclonal T cells were identified as hallmarks of this class of bispecific antibodies. Here we studied a bispecific single-chain antibody construct (referred to as 'bispecific T cell engager', BiTE) in an immunocompetent mouse model. This was possible by the use of a murine CD3-specific BiTE, and a syngeneic melanoma cell line (B16F10) expressing the human Ep-CAM target. The murine CD3-specific BiTE, called 2C11x4-7 prevented in a dose-dependent fashion the outgrowth of subcutaneously growing B16/Ep-CAM tumors with daily i.v. injections of 5 or 50 microg BiTE which was most effective. Treatment with 2C11x4-7 was effective even when it was started 10 days after tumor cell inoculation but delayed treatments showed a reduction in the number of cured animals. 2C11x4-7 was also highly active in a lung tumor colony model. When treatment was started on the day of intravenous tumor cell injection, seven out of eight animals stayed free of lung tumors, and three out of eight animals when treatment was started on day 5. Our study shows that BiTEs also have a high antitumor activity in immunocompetent mice and that there is no obvious need for costimulation of T cells by secondary agents. PMID:16187083

Schlereth, Bernd; Kleindienst, Petra; Fichtner, Iduna; Lorenczewski, Grit; Brischwein, Klaus; Lippold, Sandra; da Silva, Antonio; Locher, Mathias; Kischel, Roman; Lutterbüse, Ralf; Kufer, Peter; Baeuerle, Patrick A

2006-07-01

231

Retargeting T Cells for HER2-Positive Tumor Killing by a Bispecific Fv-Fc Antibody  

PubMed Central

To exploit the biological and pharmacological properties of immunoglobulin constant domain Fc fragment and increase the killing efficacy of T cells, a single chain variable fragment specific to CD3 was fused with Fcab (Fc antigen binding), a mutant Fc fragment with specificity against Human epidermal growth factor receptor 2 (HER2) developed by F-star. The bispecific fusion named as FcabCD3 was expressed by transient transfection in HEK-293T cells and purified by affinity chromatography. Specific cytolytic activity of retargeted T cells to kill HER2 positive SKBR3 cell line was evaluated in vitro. FcabCD3 was able to retarget T cells to kill both Herceptin insensitive Colo205-luc cell line and HER2 low expression MDA-MB-231-luc cell line. Furthermore, FcabCD3 was effective in eliminating the Colo205 tumor established on BALB/c nu/nu mice.

Wang, Lei; He, Yanran; Zhang, Ge; Ma, Juan; Liu, Changzhen; He, Wen; Wang, Wei; Han, Huamin; Boruah, Bhargavi M.; Gao, Bin

2013-01-01

232

Retargeting T cells for HER2-positive tumor killing by a bispecific Fv-Fc antibody.  

PubMed

To exploit the biological and pharmacological properties of immunoglobulin constant domain Fc fragment and increase the killing efficacy of T cells, a single chain variable fragment specific to CD3 was fused with Fcab (Fc antigen binding), a mutant Fc fragment with specificity against Human epidermal growth factor receptor 2 (HER2) developed by F-star. The bispecific fusion named as FcabCD3 was expressed by transient transfection in HEK-293T cells and purified by affinity chromatography. Specific cytolytic activity of retargeted T cells to kill HER2 positive SKBR3 cell line was evaluated in vitro. FcabCD3 was able to retarget T cells to kill both Herceptin insensitive Colo205-luc cell line and HER2 low expression MDA-MB-231-luc cell line. Furthermore, FcabCD3 was effective in eliminating the Colo205 tumor established on BALB/c nu/nu mice. PMID:24086580

Wang, Lei; He, Yanran; Zhang, Ge; Ma, Juan; Liu, Changzhen; He, Wen; Wang, Wei; Han, Huamin; Boruah, Bhargavi M; Gao, Bin

2013-01-01

233

Construction and production of an IgG-Like tetravalent bispecific antibody, IgG-single-chain Fv fusion.  

PubMed

In recent years, both laboratory and clinical studies have demonstrated that bispecific antibodies (BsAbs) may have significant potential application in cancer therapy either by targeting tumor cells with cytotoxic agents including effector cells, radionuclides, drugs, and toxins, or by simultaneously blocking two tumor-associated targets, e.g., tumor growth factors and/or their cell surface receptors. A major obstacle in the development of BsAb has been the difficulty of producing the materials in sufficient quality and quantity by traditional technologies such as the hybrid hybridoma and chemical conjugation methods. The development of recombinant BsAbs as therapeutic agents will depend heavily on the advances made in the design of the constructs (or formats) and production efficiency. Here we describe a recombinant method for the construction and production of a tetravalent IgG-like BsAb molecule, IgG-scFv fusion, in which, a single-chain Fv (scFv) antibody fragment of one antigen specificity is genetically fused to the c-terminal of a conventional IgG of a different antigen specificity. PMID:24037843

Lu, Dan; Zhu, Zhenping

2014-01-01

234

Veterinary sources of nonrodent monoclonal antibodies: interspecific and intraspecific hybridomas.  

PubMed

The generation of monoclonal antibodies from species other than rats and mice has developed slowly over the last 20 years. The advent of antibody engineering and realization of the advantages of nonmurine antibodies, in terms of their superior affinities and specificities, and their potential as components of human and veterinary therapeutics has increased their relevance recently. There have been significant advances in the development of myeloma and heteromyeloma fusion partners. This is an opportune moment to consolidate experiences of MAb production across the range of species of veterinary interest and place it into context with other developments in the field of monoclonal antibodies. The background to the development of antibodies from species other than the mouse is discussed. The species and antigens used to date are reviewed, as are the methods and results reported. A suggested protocol is provided for first attempts to exploit the huge potential of this aspect of hybridoma technology and suggestions are made for its further expansion. PMID:10952409

Groves, D J; Morris, B A

2000-06-01

235

Use of monoclonal antibodies in a radioimmunoassay for human transcortin.  

PubMed

We describe the production of monoclonal antibodies to human transcortin and their use in a radioimmunoassay (RIA). A high-affinity antibody (Ka = 4 X 10(10) L/mol) made possible a sensitive RIA for transcortin (detection limit = 0.23 ng per tube), whereas use of an antibody of moderate affinity (Ka = 5 X 10(8) L/mol) was more suitable for the routine measurement of transcortin in serum, only a 25-fold dilution of the sample being required instead of 1500-fold. The correlation was good between both RIAs (r = 0.959) and between each of the RIAs and radial immunodiffusion (r = 0.955 and 0.976 for the methods with high- and low-affinity antibody, respectively). Although monoclonal antibodies were used in the RIAs and polyclonal ones in the radial immunodiffusion procedure, similar values were obtained by all techniques. PMID:6421509

Faict, D; De Moor, P

1984-03-01

236

Understanding and circumventing resistance to anticancer monoclonal antibodies  

PubMed Central

With the widespread use of therapeutic monoclonal antibodies in the treatment of patients with cancer, resistance to these agents has become a major issue. Preclinical models of drug action or resistance have contributed to unravel the main mechanisms of resistance, involving both tumor-associated and host related factors. However our understanding of how a monoclonal antibody destroys cancer cells in a patient and why it one day stops being effective are still far from being complete. This review focuses on the available data on mechanisms of action and resistance to rituximab and includes some additional information for other monoclonal antibodies. Innovative approaches designed to overcome resistance, such as combination immunotherapy, costimulation with cytokines or growth factors are presented.

Reslan, Lina; Dalle, Stephane

2009-01-01

237

Monoclonal antibodies as disease modifying therapy in multiple sclerosis.  

PubMed

Multiple sclerosis (MS), a demyelinating disease of the central nervous system, was untreatable until the mid-1990s when beta-interferons and glatiramer acetate were introduced. These agents, while effective, were relatively nonspecific in action. Over the last 10 years, research has focused toward developing more targeted therapies for the disease. Monoclonal antibodies (mAbs) have been central to these efforts and many of the mAbs studied in MS have been singularly effective. We review here the 6 monoclonal antibodies that have been approved for MS or are in late-stage clinical trials, focusing on the drugs' efficacy and safety. Additionally, we review several monoclonal antibodies that were studied in MS but were found to be ineffective or even deleterious in this patient population. PMID:24027005

Longbrake, Erin E; Parks, Becky J; Cross, Anne H

2013-11-01

238

Intracellular forms of Drosophila topoisomerase II detected with monoclonal antibodies.  

PubMed Central

We developed monoclonal antibodies against Drosophila topoisomerase II and studied the intracellular forms and the in vivo and in vitro proteolytic degradation of the enzyme. In purified enzyme preparations polyclonal sera and monoclonal antibodies recognized several polypeptides in the 170-132 kD molecular weight range. In vivo, however, the pattern was much simpler. In Drosophila embryos, pupae, fly heads and Schneider S3 tissue culture cells topoisomerase II appeared as a single 166 kD polypeptide. In Drosophila embryos, with two monoclonal antibodies topoisomerase II appeared as a doublet composed of the 166 kD canonical form and a slightly higher molecular weight polypeptide. Topoisomerase II was shown to be present also in fly heads which are composed entirely of nonproliferative tissues. Images

Hadlaczky, G; Praznovszky, T; Sofi, J; Udvardy, A

1988-01-01

239

Monoclonal antibodies specific for African swine fever virus proteins.  

PubMed

We have obtained 60 stable hybridomas which produced immunoglobulins that recognized 12 proteins from African swine fever virus particles and African swine fever virus-infected cells. Most of the monoclonal antibodies were specific for the three major structural proteins p150, p72, and p12. The specificity of some monoclonal antibodies for the structural proteins p150 and p37 and the nonstructural proteins p220 and p60 indicated that proteins p150 and p220 are antigenically related to proteins p37 and p60. The association of some viral antigens to specific subcellular components was determined by immunofluorescence and analysis of the binding of monoclonal antibodies to infected cells. A host protein (p24) seemed to be associated with the virus particles. PMID:3882998

Sanz, A; García-Barreno, B; Nogal, M L; Viñuela, E; Enjuanes, L

1985-04-01

240

Monoclonal antibodies specific for African swine fever virus proteins.  

PubMed Central

We have obtained 60 stable hybridomas which produced immunoglobulins that recognized 12 proteins from African swine fever virus particles and African swine fever virus-infected cells. Most of the monoclonal antibodies were specific for the three major structural proteins p150, p72, and p12. The specificity of some monoclonal antibodies for the structural proteins p150 and p37 and the nonstructural proteins p220 and p60 indicated that proteins p150 and p220 are antigenically related to proteins p37 and p60. The association of some viral antigens to specific subcellular components was determined by immunofluorescence and analysis of the binding of monoclonal antibodies to infected cells. A host protein (p24) seemed to be associated with the virus particles. Images

Sanz, A; Garcia-Barreno, B; Nogal, M L; Vinuela, E; Enjuanes, L

1985-01-01

241

Humanization and simultaneous optimization of monoclonal antibody.  

PubMed

Antibody humanization is an essential technology for reducing the potential risk of immunogenicity associated with animal-derived antibodies and has been applied to a majority of the therapeutic antibodies on the market. For developing an antibody molecule as a pharmaceutical at the current biotechnology level, however, other properties also have to be considered in parallel with humanization in antibody generation and optimization. This section describes the critical properties of therapeutic antibodies that should be sufficiently qualified, including immunogenicity, binding affinity, physiochemical stability, expression in host cells and pharmacokinetics, and the basic methodologies of antibody engineering involved. By simultaneously optimizing the antibody molecule in the light of these properties, it should prove possible to shorten the research and development period necessary to identify a highly qualified clinical candidate and consequently accelerate the start of the clinical trial. PMID:24037839

Kuramochi, T; Igawa, T; Tsunoda, H; Hattori, K

2014-01-01

242

Monoclonal antibody-defined T lymphocyte subpopulations in monoclonal gammapathy of undetermined significance  

Microsoft Academic Search

Summary  We used monoclonal antibodies of the OK series to study T lymphocyte subpopulations in 55 patients with monoclonal gammapathy\\u000a of undetermined significance (MGUS) and in 40 healthy control subjects, with the aim to investigate if alterations in T lymphocyte\\u000a subpopulations occur also in MGUS. Mean OKT3+ and OKT8+ cell counts were higher (p<0.01 and p<0.001, respectively) and the mean OKT4\\/OKT8

Ettore Cunietti; Massimo Galli; Cosimo Ottomano; Cristina Negri; Laura Soldini; Luisa Scapellato; Giancarlo Sensalari

1987-01-01

243

Application of Quantitative Pharmacology in Development of Therapeutic Monoclonal Antibodies  

PubMed Central

The advancement of therapeutic monoclonal antibodies during various stages of the drug development process can be effectively streamlined when appropriate translational strategies are applied. Design of successful translational strategies for development of monoclonal antibodies should allow for understanding of the dose– and concentration–response relationships with respect to both beneficial and toxic effects from early phases of drug development. Evaluation of relevant biomarkers during early stages of drug development should facilitate the successful design of safe and effective dosing strategies. Moreover, application of quantitative pharmacology is critical for translation of exposure–response relationships early on.

Funelas, Cherryl; Suria, Hamza

2010-01-01

244

[ADLib system for rapid and flexible design of monoclonal antibodies].  

PubMed

Monoclonal antibodies (MAbs) have been utilized as research tools, as diagnostic reagents, and for antibody medicine. The preparation of MAbs involves laborious processes and normally takes months. Here we describe an ex vivo B cell-based antibody display system called the ADLib (Autonomously Diversifying Library) system, which enables us to select chicken B cell clones producing antibody against antigens of interest in a couple of weeks. The ADLib system is applicable to self- or highly conserved antigens, polysaccharide chains, peptides, and haptens. PMID:17202787

Ohta, Kunihiro; Seo, Hidetaka

2007-01-01

245

Specific Recognition of Low Density Lipoprotein subspecies from Hypertriglyceridemic Subjects by a Monoclonal Antibody.  

National Technical Information Service (NTIS)

The authors have tested the reactivity of an anti-apoliprotein B monoclonal antibody (IVA5) with plasma low density lipoprotein (LSL) from patients with hypertriglyceridemia (HTG). Keywords: Monoclonal antibodies; Low density Lipoprotein; Reprints.

R. Cubicciotti R. M. Krauss A. E. Karu

1984-01-01

246

MONOCLONAL ANTIBODIES IDENTIFY CONSERVED EPITOPES ON THE POLYHEDRIN OF 'HELIOTHIS ZEA' NUCLEAR POLYHEDROSIS VIRUS  

EPA Science Inventory

Recent advances in monoclonal antibody techniques have provided an opportunity to simplify the procedures of serological identification of microorganisms. Because monoclonal antibodies are raised against individual antigenic determinants (epitopes), they can be used to screen wit...

247

Hybridomas and Monoclonal Antibodies Therefrom Having Specific Reactivity Toward Heavy Chain Immunoglobulin from Catfish.  

National Technical Information Service (NTIS)

A hybridoma cell line which produces a monoclonal antibody with reactivity directed specifically to catfish immunoglobulin heavy chain having a molecular weight of 70,000 daltons was discovered. The hybridoma monoclonal antibody is specific for immunoglob...

P. H. Klesius

1989-01-01

248

Antibody discovery: sourcing of monoclonal antibody variable domains.  

PubMed

Historically, antibody variable domains for therapeutic antibodies have been sourced primarily from the mouse IgG repertoire, and typically either chimerized or humanized. More recently, human antibodies from transgenic mice producing human IgG, phage display libraries, and directly from human B lymphocytes have been used more broadly as sources of antibody variable domains for therapeutic antibodies. Of the total 36 antibodies approved by major maket regulatory agencies, the variable domain sequences of 26 originate from the mouse. Of these, four are marketed as murine antibodies (of which one is a mouse-rat hybrid IgG antibody), six are mouse-human chimeric antibodies, and 16 are humanized. Ten marketed antibodies have originated from human antibody genes, three isolated from phage libraries of human antibody genes and seven from transgenic mice producing human antibodies. Five antibodies currently in clinical trials have been sourced from camelids, as well as two from non-human primates, one from rat, and one from rabbit. Additional sources of antibody variable domains that may soon find their way into the clinic are potential antibodies from sharks and chickens. Finally, the various methods for retrieval of antibodies from humans, mouse and other sources, including various display technologies and amplification directly from B cells, are described. PMID:24168292

Strohl, William R

2014-03-01

249

Therapeutic window of MuS110, a single-chain antibody construct bispecific for murine EpCAM and murine CD3.  

PubMed

EpCAM (CD326) is one of the most frequently and highly expressed tumor-associated antigens known and recently has also been found on cancer stem cells derived from human breast, colon, prostate, and pancreas tumors. However, like many other tumor-associated antigens used for antibody-based immunotherapeutic approaches, EpCAM is expressed on normal tissues including epithelia of pancreas, colon, lung, bile ducts, and breast. To assess the therapeutic window of an EpCAM/CD3-bispecific single-chain antibody construct of the bispecific T-cell engager (BiTE) class, we constructed murine surrogate of MT110 (muS110) from single-chain antibodies specific for murine EpCAM and CD3 antigens. Immunhistochemical analysis showed that, with minor differences, the expression of EpCAM protein on a large variety of tissues from man and mouse was similar with respect to distribution and level. MuS110 exhibited significant antitumor activity at as low as 5 microg/kg in both syngeneic 4T1 orthotopic breast cancer and CT-26 lung cancer mouse models. Dosing of muS110 for several weeks up to 400 microg/kg by intraanimal dose escalation was still tolerated, indicating existence of a significant therapeutic window for an EpCAM-specific BiTE antibody in mice. MuS110 was found to have similar in vitro characteristics and in vivo antitumor activity as MT110, a human EpCAM/human CD3-bispecific BiTE antibody that currently is in formal preclinical development. PMID:18172306

Amann, Maria; Brischwein, Klaus; Lutterbuese, Petra; Parr, Larissa; Petersen, Laetitia; Lorenczewski, Grit; Krinner, Eva; Bruckmeier, Sandra; Lippold, Sandra; Kischel, Roman; Lutterbuese, Ralf; Kufer, Peter; Baeuerle, Patrick A; Schlereth, Bernd

2008-01-01

250

Radioiodination of monoclonal antibody for prostate specific antigen  

Microsoft Academic Search

Prostate specific antigen (PSA) is a reliable biochemical marker used in screening for prostate carcinoma. Immunoradiometric assays (IRMA) are generally used for the estimation of total\\/free PSA in serum samples. Radiolabeled antibody, an important reagent of IRMA was prepared and characterized using an in-house anti-PSA monoclonal antibody (Mab), Mab-2S. Mab-2S was radiolabeled with 125I and characterized for immunoreactivity and radiochemical

K. Bapat; A. Korde; M. Zaidi; A. Mukherjee; M. R. A. Pillai; M. Venkatesh

2002-01-01

251

Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies  

NASA Astrophysics Data System (ADS)

Mixtures of certain monoclonal antibodies appear to bind human chorionic gonadotropin in a ``cooperative'' fashion because they form circular complexes with the hormone. Experiments illustrate how this property might be exploited to develop very sensitive immunoassays for human chorionic gonadotropin or any other antigen. Since the assays are not based on competitive inhibition between radiolabeled and unlabeled antigen, they are much more sensitive than a traditional radioimmunoassay in which either one of the same antibodies is used alone.

Ehrlich, Paul H.; Moyle, William R.

1983-07-01

252

Monoclonal antibody against the glutaraldehyde-conjugated polyamine, spermine  

Microsoft Academic Search

We developed a mouse monoclonal antibody (ASPM-29, mAb) against spermine (Spm) conjugated to human serum albumin (HSA) using glutaraldehyde-sodium borohydride, for applications in immunocytochemistry (ICC). The antibody specificity was evaluated by an enzyme-linked immunosorbent assay (ELISA) binding test, simulating the ICC of tissue sections. ASPM-29 showed an almost equal immunoreactivity to Spm and spermidine (Spd) but no reactivity to any

Kunio Fujiwara; Yukinobu Masuyama

1995-01-01

253

Production and Characterization of Monoclonal Antibodies Against Aflatoxin G 1  

Microsoft Academic Search

Monoclonal antibodies against ochratoxin B (OTB) were generated by immunizing Balb\\/c mice with OTB conjugated to keyhole limpet hemocyanin (KLH) via carbodiimide reactions with CHMC and EDAC. A stable hybridoma cell line 2F1.E10 was produced by fusion of murine splenocytes and myeloma cells. The obtained antibodies were characterized using an indirect competitive ELISA. The detection limit was calculated (27 ±

Jinyang Zhang; Peiwu Li; Wen Zhang; Qi Zhang; Xiaoxia Ding; Xiaomei Chen; Wenhua Wu; Xianzhong Zhang

2009-01-01

254

The rapid quantification and detection of nitrifying bacteria by using monoclonal antibody method  

Microsoft Academic Search

Monoclonal antibodies against the two kinds of nitrifying bacteria Nitrosomonas europaea (IFO14298) and Nitrobacter winogradskyi (IFO14297) were raised and isotypes of these monoclonal antibodies, IgM and IgG1, were successfully obtained. Cross reactivities of these monoclonal antibodies against various kinds of representative heterotrophic bacteria turned out to be relatively low by competitive ELISA. In contrast, these monoclonal antibodies were very specific

H. Ikuta; N. Nod; Y. Ebie; A. Hirata; S. Tsuneda; M. Matsumura; Y. Inamori

2000-01-01

255

Method for imaging breast tumors using labeled monoclonal anti-human breast cancer antibodies  

US Patent & Trademark Office Database

Hybridomas producing monoclonal antibodies suitable for imaging and diagnosis of human breast tumors and such monoclonal antibodies are claimed. The monoclonals are characterized by breast tumor binding range, breast cancer cell line range, and selectivity. Immunoimaging agents comprising the monoclonal antibody and a detectable label, either directly or indirectly conjugated to the antibody are claimed. Methods for imaging breast tumors using the immunoimaging agents are described and claimed.

1990-07-03

256

Rescue of Impaired NK Cell Activity in Hodgkin Lymphoma With Bispecific Antibodies In Vitro and in Patients  

PubMed Central

Natural killer (NK) cells represent a key component of the innate immune system against cancer. Nevertheless, malignant diseases arise in immunocompetent individuals despite tumor immunosurveillance. Hodgkin lymphoma (HL) is characterized by CD30+ tumor cells and a massive infiltration of immune effector cells in affected lymph nodes. The latter obviously fail to eliminate the malignant cell population. Here, we tested for functional NK cell defects in HL and suggest an improvement of NK function by therapeutic means. We demonstrate that peripheral NK cells (pNK) from patients with HL fail to eliminate HL cell lines in ex vivo killing assays. Impaired NK cell function correlated with elevated serum levels of soluble ligands for NK cell receptors NKp30 (BAG6/BAT3) and NKG2D (MICA), factors known to constrict NK cell function. In vitro, NK cell cytotoxicity could be restored by an NKG2D/NKp30-independent bispecific antibody construct (CD30xCD16A). It artificially links the tumor receptor CD30 with the cytotoxicity NK cell receptor CD16A. Moreover, we observed that NK cells from patients treated with this construct were generally activated and displayed a restored cytotoxicity against HL target cells. These data suggest that reversible suppression of NK cell activity contributes to immune evasion in HL and can be antagonized therapeutically.

Reiners, Katrin S.; Kessler, Jorg; Sauer, Maike; Rothe, Achim; Hansen, Hinrich P.; Reusch, Uwe; Hucke, Christian; Kohl, Ulrike; Durkop, Horst; Engert, Andreas; von Strandmann, Elke Pogge

2013-01-01

257

Rescue of impaired NK cell activity in hodgkin lymphoma with bispecific antibodies in vitro and in patients.  

PubMed

Natural killer (NK) cells represent a key component of the innate immune system against cancer. Nevertheless, malignant diseases arise in immunocompetent individuals despite tumor immunosurveillance. Hodgkin lymphoma (HL) is characterized by CD30(+) tumor cells and a massive infiltration of immune effector cells in affected lymph nodes. The latter obviously fail to eliminate the malignant cell population. Here, we tested for functional NK cell defects in HL and suggest an improvement of NK function by therapeutic means. We demonstrate that peripheral NK cells (pNK) from patients with HL fail to eliminate HL cell lines in ex vivo killing assays. Impaired NK cell function correlated with elevated serum levels of soluble ligands for NK cell receptors NKp30 (BAG6/BAT3) and NKG2D (MICA), factors known to constrict NK cell function. In vitro, NK cell cytotoxicity could be restored by an NKG2D/NKp30-independent bispecific antibody construct (CD30xCD16A). It artificially links the tumor receptor CD30 with the cytotoxicity NK cell receptor CD16A. Moreover, we observed that NK cells from patients treated with this construct were generally activated and displayed a restored cytotoxicity against HL target cells. These data suggest that reversible suppression of NK cell activity contributes to immune evasion in HL and can be antagonized therapeutically. PMID:23459515

Reiners, Katrin S; Kessler, Jörg; Sauer, Maike; Rothe, Achim; Hansen, Hinrich P; Reusch, Uwe; Hucke, Christian; Köhl, Ulrike; Dürkop, Horst; Engert, Andreas; von Strandmann, Elke Pogge

2013-04-01

258

Effect of small-molecule-binding affinity on tumor uptake in vivo: a systematic study using a pretargeted bispecific antibody.  

PubMed

Small-molecule ligands specific for tumor-associated surface receptors have wide applications in cancer diagnosis and therapy. Achieving high-affinity binding to the desired target is important for improving detection limits and for increasing therapeutic efficacy. However, the affinity required for maximal binding and retention remains unknown. Here, we present a systematic study of the effect of small-molecule affinity on tumor uptake in vivo with affinities spanning a range of three orders of magnitude. A pretargeted bispecific antibody with different binding affinities to different DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid)-based small molecules is used as a receptor proxy. In this particular system targeting carcinoembryonic antigen, a small-molecule-binding affinity of 400 pmol/L was sufficient to achieve maximal tumor targeting, and an improvement in affinity to 10 pmol/L showed no significant improvement in tumor uptake at 24 hours postinjection. We derive a simple mathematical model of tumor targeting using measurable parameters that correlates well with experimental observations. We use relations derived from the model to develop design criteria for the future development of small-molecule agents for targeted cancer therapeutics. PMID:22491799

Orcutt, Kelly Davis; Rhoden, John J; Ruiz-Yi, Benjamin; Frangioni, John V; Wittrup, K Dane

2012-06-01

259

Limitations of radiolabeled monoclonal antibodies for localization of human neoplasms  

SciTech Connect

Tumor-associated monoclonal antibodies were radiolabeled with /sup 125/I and /sup 131/I and given i.v. in pairs to 19 patients 1-26 days prior to surgical excision of primary and metastatic breast, ovarian, and gastrointestinal tumors. For individual patients each monoclonal antibody was designated as specific or nonspecific according to prior immunoperoxidase staining results on the appropriate target neoplastic tissues. Quantitation of antibody uptake was performed on resected normal and neoplastic tissues. Although good tumor:non-tumor ratios were obtained with the specific antibodies (maximal tumor:blood ratio, 35.8:1 at 12 days postadministration), the absolute amount of radiolabel detected in tumors was small (mean value of 0.015% of total injected amount per g of tumor occurring 1 day postadministration). Furthermore, both specific and nonspecific antibodies accumulated in normal lymph nodes to a significant extent (mean value of 0.0026% of total injected amount per g of tissue occurring 1 day postadministration). Knowledge of such data is essential prior to considering therapeutic uses of radiolabeled monoclonal antibodies.

Epenetos, A.A.; Snook, D.; Durbin, H.; Johnson, P.M.; Taylor-Papadimitriou, J.

1986-06-01

260

Identification of Aeromonas hydrophila infection with specific monoclonal antibodies  

Microsoft Academic Search

Whole cell of Aeromonas hydrophila 1234 was used for immunization to produce monoclonal antibodies (MAbs). Three different groups of MAbs specific to Aeromonas were obtained. The first group of MAbs demonstrated high specificity and bound to the A. hydrophila 1234 only but did not bind to the other two A. hydrophila isolates. This group of MAbs bound to a series

Siwaporn Longyant; Kriengkrai Prahkarnkaeo; Vithaya Meevoothisom; Sirirat Rengpipat; Sombat Rukpratanporn; Parin Chaivisuthangkura

261

Serological study of yeast killer toxins by monoclonal antibodies  

Microsoft Academic Search

Yeast killer toxins coded by determined and undetermined killer plasmids or presumptive nuclear gene(s) in various genera (Saccharomyces, Kluyveromyces, Pichia and Candida) have been serologically investigated by a monoclonal antibody (KT4), produced against the yeast killer toxin of Pichia (Hansenula) anomala UCSC 25F. Double immunodiffusion with the killer toxins as antigens and indirect immunofluorescence on whole cells of the corresponding

Luciano Polonelli; Stefania Manzara; Stefania Conti; Giuseppe Dettori; Giulia Morace; Carlo Chezzi

1989-01-01

262

EGFR targeting therapies: Monoclonal antibodies versus tyrosine kinase inhibitors  

Microsoft Academic Search

Current development of targeted therapy in oncology is particularly active and concerns principally two types of agents which are monoclonal antibodies (Mabs) and tyrosine kinase inhibitors (TKIs). Epidermal growth factor receptor (EGFR) signaling pathways play a key role in the regulation of cell proliferation, survival and differentiation. Consequently, EGFR is one of the most-studied ligand–receptor systems and specific EGFR inhibition

Olivier Dassonville; Alexandre Bozec; Jean Louis Fischel; Gerard Milano

2007-01-01

263

Antigenic analysis of West Nile virus strains using monoclonal antibodies  

Microsoft Academic Search

Summary Seventeen monoclonal antibodies (MAbs) were prepared against the flavivirus West Nile strain H442. While the majority of these were specific for the major envelope protein, MAbs directed against the NS1 and ns4a nonstructural proteins were also identified. The MAbs were tested by indirect immunofluorescence against 16 southern African West Nile (WN) isolates, representative strains from the two main WN

Terry G. Besselaar; N. K. Blackburn

1988-01-01

264

Indium-111 labeled anti-melanoma monoclonal antibodies  

DOEpatents

A monoclonal antibody to a high molecular weight melanoma-associated antigen was chelated and radiolabeled with indium-111. This material shows high affinity for melanoma and thus can be used in the detection, localization and imaging of melanoma. 1 figure.

Srivastava, S.C.; Fawwaz, R.A.; Ferrone, S.

1984-04-30

265

Boronated Dextran-Monoclonal Antibody Conjugates for NCT.  

National Technical Information Service (NTIS)

Monoclonal antibodies (MoAbs) to tumor-associated antigens are attractive carriers for concentrating sup 10 B in target tissues. Since coupling of sup 10 B compounds directly to MoAb in amounts sufficient for radiotherapy appears to lead to inactive compl...

J. J. Elmore D. C. Borg D. Gabel R. G. Fairchild M. Temponi

1985-01-01

266

A panel immunoblot using co-incubated monoclonal antibodies for identification of melanoma cells  

Microsoft Academic Search

Antigen expression in melanoma is heterogeneous. Immunophenotyping using a panel of monoclonal antibodies may facilitate immunotherapy. An immunoblot procedure was developed to detect antigens in melanoma cells. Numerous monoclonal antibodies were tested to determine if (1) antigens were detected after transfer to membranes, (2) single bands or discrete multiple bands were obtained, (3) co-incubation of multiple monoclonal antibodies had no

Rong Wang; Linda J. Dworak; Michael J. Lacy

2001-01-01

267

Platelet Imaging in Man using Antiplatelet Monoclonal Antibodies.  

PubMed

Radiolabeled platelets have been used in experimental imaging studies using the gamma camera to localize and quantify platelet accumulation. Conventional radiolabelling methods have been laborious, requiring specialized expertise and facilities. Recent advances in biotechnology have provided a number of murine monoclonal antibodies for the selective radiolabelling of circulating cells, cellular components and proteins. Most of these antibodies have been used for the in vitro study of platelet membrane structures. Relatively few antibodies have been administered to humans for in vivo imaging. Nevertheless, the use of monoclonal antibodies as a means of in vivo radiolabelling of platelets following a single intravenous injection represents a significant advance over the established techniques. Clinical studies currently underway have demonstrated that antibodies specific for platelets have the potential for replacing the conventional methods of radiolabelling and can result in images of outstanding quality. This paper reviews the current experimental and clinical studies using antibodies specific for platelets. In the main, these antibodies bind to the platelet membrane glycoprotein complexes IIb/IIIa or Ia/IIa. Recently, however, an antibody binding to P-selectin, and therefore specific to activated platelets, has been introduced and represents an interesting new development. The future application of this technology based on either humanized or synthetic molecules recognizing platelet antigens is likely to become routinely accepted for the study of platelet deposition in experimental settings and for thrombus imaging in clinical practice. PMID:21043856

Perkins, A C; Lonsdale, R J

1993-01-01

268

Hexavalent bispecific antibodies represent a new class of anticancer therapeutics: 1. Properties of anti-CD20/CD22 antibodies in lymphoma  

PubMed Central

The dock and lock (DNL) method is a new technology for generating multivalent antibodies. Here, we report in vitro and in vivo characterizations of 20-22 and 22-20, a pair of humanized hexavalent anti-CD20/22 bispecific antibodies (bsAbs) derived from veltuzumab (v-mab) and epratuzumab (e-mab). The 22-20 was made by site-specific conjugation of e-mab to 4 Fabs of v-mab; 20-22 is of the opposite configuration, composing v-mab and 4 Fabs of e-mab. Each bsAb translocates both CD22 and CD20 into lipid rafts, induces apoptosis and growth inhibition without second-antibody crosslinking, and is significantly more potent in killing lymphoma cells in vitro than their parental antibodies. Although both bsAbs triggered antibody-dependent cellular toxicity, neither displayed complement-dependent cytotoxicity. Intriguingly, 22-20 and 20-22 killed human lymphoma cells in preference to normal B cells ex vivo, whereas the parental v-mab depleted malignant and normal B cells equally. In vivo studies in Daudi tumors revealed 20-22, despite having a shorter serum half-life, had antitumor efficacy comparable with equimolar v-mab; 22-20 was less potent than 20-22 but more effective than e-mab and control bsAbs. These results indicate multiple advantages of hexavalent anti-CD20/22 bsAbs over the individual parental antibodies and suggest that these may represent a new class of cancer therapeutics.

Cardillo, Thomas M.; Stein, Rhona; Chang, Chien-Hsing

2009-01-01

269

Purification of human monoclonal antibodies and their fragments.  

PubMed

This chapter summarizes the most common chromatographic mAb and mAb fragment purification methods, starting by elucidating the relevant properties of the compounds and introducing the various chromatography modes that are available and useful for this application. A focus is put on the capture step affinity and ion exchange chromatography. Aspects of scalability play an important role in judging the suitability of the methods. The chapter introduces also analytical chromatographic methods that can be utilized for quantification and purity control of the product. In the case of mAbs, for most purposes the purity obtained using an affinity capture step is sufficient. Polishing steps are required if material of particularly high purity needs to be generated. For mAb fragments, affinity chromatography is not yet fully established, and the capture step potentially may not provide material of high purity. Therefore, the available polishing techniques are touched upon briefly. In the case of mAb isoform and bispecific antibody purification, countercurrent chromatography techniques have been proven to be very useful and a part of this chapter has been dedicated to them, paying tribute to the rising interest in these antibody formats in research and industry. PMID:24037849

Müller-Späth, Thomas; Morbidelli, Massimo

2014-01-01

270

Monoclonal antibodies produced by muscle after plasmid injection and electroporation.  

PubMed

Antibodies are useful for the treatment of a variety of diseases. We here demonstrate that mouse muscle produced monoclonal antibodies (mAb) after a single injection of immunoglobulin genes as plasmid DNA. In vivo electroporation of muscle greatly enhanced antibody production. For chimeric antibodies, levels of 50-200 ng mAb/ml serum were obtained but levels declined after 7-14 days due to an immune response against the xenogeneic parts of the antibody. By contrast, fully mouse antibodies persisted in serum for at least 7 months. mAb produced by the muscle had correct structure, specificity, and biological effector functions. The findings were extended to a larger animal, the sheep, in which mAb serum levels of 30-50 ng/ml were obtained. Sustained levels of serum mAb, induced by single injection of Ig genes and electroporation of muscle cells, may offer significant advantages in the treatment of human diseases. PMID:15006599

Tjelle, Torunn Elisabeth; Corthay, Alexandre; Lunde, Elin; Sandlie, Inger; Michaelsen, Terje E; Mathiesen, Iacob; Bogen, Bjarne

2004-03-01

271

Monoclonal antibodies against farmer's lung antigens having specific binding to IgG antibodies.  

PubMed

Hypersensitivity pneumonitis resulting from environmental exposure to Saccharopolyspora rectivirgula (Micropolyspora faeni) among farmers has been well recognized. The diagnosis of the disease depends on demonstration of circulating antibodies against S. rectivirgula. However, dependable pure antigens are not available for serodiagnosis. In the present study we have employed hybridoma technology to obtain monoclonal antibodies against S. rectivirgula antigens. These monoclonal antibodies were employed to purify antigens through affinity chromatography. When tested in ELISA, high levels of antibodies were demonstrated against these antigens in farmer's lung patient sera compared to exposed but asymptomatic individuals from the same household. In Western blots patient sera reacted with components of crude antigens with molecular masses of 28, 35, 60, 65 and 68 kD and 4 components above 100 kD, while the monoclonal antibodies reacted only with the 60-kD protein. These purified antigens can be used as reliable reagents in the specific diagnosis of farmer's lung disease. PMID:8400887

Kumar, A; Elms, N; Kurup, V P

1993-01-01

272

Library of monoclonal antibodies against brush border membrane epithelial antigens  

SciTech Connect

A purified fraction of proximal tubule brush border membranes (BBM) was prepared from dog kidney and used to immunize mice. The standard technique of hybridoma production was followed as described by Kohler and Milstein. Production of antibodies was detected by indirect immunofluorescence on dog kidney slices and by immunodot against the purified fraction on nitrocellulose. Five hybrids exhibited anti BBM activity. These were cloned twice and yielded stable cell lines producing IgG type monoclonal antibodies against BBM. They were designated A/sub 1/, C/sub 7/, D/sub 3/, D/sub 7/ and H/sub 4/. As a family these five monoclonals have broad tissue specificity, i.e. positive staining of the surface mucosa of intestinal kidney proximal tubules. D/sub 3/ exhibits even broader specificity for epithelium reacting with bile canaliculi and choroid plexus. The authors have verified that at least 4/5 antibodies are directed against BBM protein as revealed by immunoprecipitation of solubilized BBM and detected by Coomassie blue staining or autoradiography of lactoperoxidase labelled BBM. Most interestingly all antibodies bind to the surface of LL CPK/sub 1/ cells, a continuous pig kidney cell line of undefined origin but exhibiting many characteristics of proximal tubule cells. The library of monoclonal antibodies obtained provide important probes with which to study membrane biogenesis and polarization in epithelial cells.

Behar, M.; Katz, A.; Silverman, M.

1986-03-01

273

High efficiency iodination of monoclonal antibodies for radiotherapy  

SciTech Connect

A technique for labeling monoclonal antibodies (MoAb) with large activities of radioiodine using the reagent N-bromosuccinimide is described. This technique has been used to label three tumor-associated antibodies with an average labeling efficiency of greater than 90%. No significant damage to the antibody could be detected by enzyme linked immunosorbent assay (ELISA), performed immediately after the labeling procedure. Because high radiochemical purity can be achieved without the need postlabeling purification, the procedure is rapid and results in very low radiation doses to staff.

Mather, S.J.; Ward, B.G.

1987-06-01

274

Generation of Monoclonal Antibodies against Highly Conserved Antigens  

PubMed Central

Background Therapeutic antibody development is one of the fastest growing areas of the pharmaceutical industry. Generating high-quality monoclonal antibodies against a given therapeutic target is very crucial for the success of the drug development. However, due to immune tolerance, some proteins that are highly conserved between mice and humans are not very immunogenic in mice, making it difficult to generate antibodies using a conventional approach. Methodology/Principal Findings In this report, the impaired immune tolerance of NZB/W mice was exploited to generate monoclonal antibodies against highly conserved or self-antigens. Using two highly conserved human antigens (MIF and HMGB1) and one mouse self-antigen (TNF-alpha) as examples, we demonstrate here that multiple clones of high affinity, highly specific antibodies with desired biological activities can be generated, using the NZB/W mouse as the immunization host and a T cell-specific tag fused to a recombinant antigen to stimulate the immune system. Conclusions/Significance We developed an efficient and universal method for generating surrogate or therapeutic antibodies against “difficult antigens” to facilitate the development of therapeutic antibodies.

Zhou, Hongzhe; Wang, Yunbo; Wang, Wei; Jia, Junying; Li, Yuan; Wang, Qiyu; Wu, Yanfang; Tang, Jie

2009-01-01

275

Treating multiple sclerosis with monoclonal antibodies: a 2013 update.  

PubMed

The third part of this in-depth review series on the treatment of multiple sclerosis (MS) with monoclonal antibodies covers the years 2010-2012. The natalizumab section gives a progressive multifocal leukoencephalopathy update, focusing on clinically relevant aspects. Furthermore, it outlines problems around natalizumab cessation and current evidence on therapeutic strategies thereafter. Finally, it reviews evidence on Janus-faced modes of natalizumab action besides anti-inflammatory effects, including proinflammatory effects. The section on alemtuzumab critically analyzes recent Phase III results and discusses which patients might be best suited for alemtuzumab treatment, and reviews the long-term immunological impact of this anti-CD52 antibody. The daclizumab section critically summarizes results from the Phase IIb SELECT/SELECTION trial and introduces the Phase III program. The section on anti-CD20 antibodies reviews Phase II results on ocrelizumab and ofatumumab, and discusses current perspectives of these antibodies for MS therapy. Promising recent Phase II results on the anti-IL-17A antibody secukinumab (AIN457) are outlined and a short update on tabalumab (LY2127399) is given. Other highlighted antibodies currently being tested in MS patients include GNbAC1, BIIB033, MOR103 and MEDI-551. Finally, the authors give an update on the role monoclonal antibodies could play in the therapeutic armamentarium for MS in the medium term. PMID:23448220

Deiß, Annika; Brecht, Isabel; Haarmann, Axel; Buttmann, Mathias

2013-03-01

276

Proteomic identification of monoclonal antibodies from serum.  

PubMed

Characterizing the in vivo dynamics of the polyclonal antibody repertoire in serum, such as that which might arise in response to stimulation with an antigen, is difficult due to the presence of many highly similar immunoglobulin proteins, each specified by distinct B lymphocytes. These challenges have precluded the use of conventional mass spectrometry for antibody identification based on peptide mass spectral matches to a genomic reference database. Recently, progress has been made using bottom-up analysis of serum antibodies by nanoflow liquid chromatography/high-resolution tandem mass spectrometry combined with a sample-specific antibody sequence database generated by high-throughput sequencing of individual B cell immunoglobulin variable domains (V genes). Here, we describe how intrinsic features of antibody primary structure, most notably the interspersed segments of variable and conserved amino acid sequences, generate recurring patterns in the corresponding peptide mass spectra of V gene peptides, greatly complicating the assignment of correct sequences to mass spectral data. We show that the standard method of decoy-based error modeling fails to account for the error introduced by these highly similar sequences, leading to a significant underestimation of the false discovery rate. Because of these effects, antibody-derived peptide mass spectra require increased stringency in their interpretation. The use of filters based on the mean precursor ion mass accuracy of peptide-spectrum matches is shown to be particularly effective in distinguishing between "true" and "false" identifications. These findings highlight important caveats associated with the use of standard database search and error-modeling methods with nonstandard data sets and custom sequence databases. PMID:24684310

Boutz, Daniel R; Horton, Andrew P; Wine, Yariv; Lavinder, Jason J; Georgiou, George; Marcotte, Edward M

2014-05-20

277

Cloned transgenic farm animals produce a bispecific antibody for T cell-mediated tumor cell killing  

Microsoft Academic Search

Complex recombinant antibody fragments for modulation of immune function such as tumor cell destruction have emerged at a rapid pace and diverse anticancer strategies are being developed to benefit patients. Despite improvements in molecule design and expression systems, the quantity and stability, e.g., of single-chain antibodies produced in cell culture, is often insufficient for treatment of human disease, and the

Ludger Grosse-Hovest; Sigrid Müller; Rosa Minoia; Eckhard Wolf; Valeri Zakhartchenko; Hendrik Wenigerkind; Caroline Lassnig; Urban Besenfelder; Mathias Müller; Simon D. Lytton; Gundram Jung; Gottfried Brem

2004-01-01

278

Comparison of rabbit monoclonal and mouse monoclonal antibodies in immunohistochemistry in canine tissues.  

PubMed

Rabbit monoclonal (RM) antibodies appear to have higher affinity for antigens than mouse monoclonal (MM) antibodies. However, RM antibodies have not been used in veterinary diagnostic immunohistochemistry. The authors compared reactivities of RM and MM antibodies on formalin-fixed, paraffin-embedded canine tissues, targeting 11 different antigens: CD3, CD79a, calcitonin, calretinin, chromogranin A, COX-2, estrogen receptor, Ki67, progesterone receptor, synaptophysin, and vimentin. Paraffin-embedded tissue sections were processed by 1 of 2 antigen-retrieval methods: 1) proteinase K digestion or 2) steam heat in citrate buffer. An additional set of slides did not receive antigen retrieval. Immunostaining was performed using an automated stainer, and scores were assigned to the different dilutions and antigen-retrieval methods on the basis of staining intensity and number of positive cells. Steam heat was usually the best antigen-retrieval method. The optimal dilution for each antibody was that which resulted in the highest specific staining and the lowest nonspecific (background) staining. The RM or MM antibodies yielded a specific reaction for all antigens examined except calretinin. The RM and MM antibodies yielded a specific reaction for 4 antigens only: COX-2, Ki67, synaptophysin, and vimentin. Three antigens (CD3, chromogranin A, and progesterone receptor) were detected only with RM antibodies, whereas the other 3 (CD79a, calcitonin, estrogen receptor) were detected only with MM antibodies. The results of this study differed from those reported for human tissues by the manufacturers of the antibodies. These results emphasize that, regardless of manufacturers' recommendations, each antibody must be individually standardized and validated before routine use in canine tissues. PMID:16130992

Vilches-Moure, José G; Ramos-Vara, José A

2005-07-01

279

Ongoing Development of Monoclonal Antibodies and Antibody Drug-Conjugates in Lymphoma  

Microsoft Academic Search

Rituximab, a monoclonal antibody (mAb) directed against CD20, has changed practices in the treatment of patients with B-cell\\u000a lymphoma. The large success of rituximab has contributed to validate immunotherapy with monoclonal antibodies as a valuable\\u000a strategy in lymphoma. Recently, better-engineered anti-CD20-mAbs have been designed to improve efficacy and safety. Also,\\u000a new antibodies targeting other lymphoma subtypes including T-cell lymphoma and

Eileen M. Boyle; Franck Morschhauser

280

Monoclonal antibodies identify a group of nuclear pore complex glycoproteins  

PubMed Central

Using monoclonal antibodies we identified a group of eight polypeptides of rat liver nuclear envelopes that have common epitopes. Most or all of these proteins are structurally distinct, as shown by tryptic peptide mapping and analysis with polyclonal antibodies. While these polypeptides are relatively tightly bound to nuclear membranes, only one is an integral membrane protein. The eight antigens cofractionate with the nuclear pore complex under various conditions of ionic strength and detergent. It can be seen by immunofluorescence microscopy that the monoclonal antibodies reacting with these antigens stain the nuclear surface of interphase cells in a finely punctate pattern. When the nuclear envelope is disassembled and subsequently reformed during mitosis, the proteins are reversibly dispersed throughout the cytoplasm in the form of minute foci. By EM immunogold localization on isolated nuclear envelopes, the monoclonal antibodies label exclusively the nuclear pore complex, at both its nucleoplasmic and cytoplasmic margins. Considered together, our biochemical and localization data indicate that the eight nuclear envelope polypeptides are pore complex components. As shown in the accompanying paper (Holt, G. D., C. M. Snow, A. Senior, R. S. Haltiwanger, L. Gerace, and G. W. Hart, J. Cell Biol., 104:1157-1164) these eight polypeptides contain a novel form of glycosylation, O-linked N-acetylglucosamine. The relative abundance and disposition of these O-linked glycoproteins in the pore complex are consistent with their having a role in nucleocytoplasmic transport.

1987-01-01

281

The Use of Monoclonal Antibodies in Human Prion Disease  

NASA Astrophysics Data System (ADS)

Detection of PrP and its pathological isoform(s) is the key to understanding the etiology and pathogenesis of transmissible spongiform encephalopathy. There is ample evidence that PrP isoforms constitute a major component of an unknown and perhaps unconventional infectious agent. An etiological relationship between human and zoonotic transmissible spongiform encephalopathies may be revealed with monoclonal antibodies. Knowledge of the conformational transition rendering a nonpathogenic, almost ubiquitous cellular protein into a pathogenic one is crucial to defining pathomechanisms. The stepwise or even continuous formation of pathogenic molecules can be monitored. Any improvement in the early diagnosis could help to conceive new therapeutic measures which are not currently available. Determination of PrP isoforms in tissue, cells, or body fluids may be of prognostic value. Many experimental approaches in molecular medicine and molecular biology of the prion protein already rely on monoclonal antibodies. Recombinant antibodies such as the single-chain Fv may soon replace traditional hybridoma techniques. Binding affinity can easily be manipulated by a number of techniques, including in vitro mutagenesis - a step which could never be carried out using the traditional hybridoma technology. Monoclonal antibodies are and will remain an essential support for ongoing research on the prion protein in general and on the unconventional infectious prions.

Bodemer, Walter

282

Stimuli-responsive magnetic nanoparticles for monoclonal antibody purification.  

PubMed

Monoclonal antibodies (mAbs) are important therapeutic proteins. One of the challenges facing large-scale production of monoclonal antibodies is the capacity bottleneck in downstream processing, which can be circumvented by using magnetic stimuli-responsive polymer nanoparticles. In this work, stimuli-responsive magnetic particles composed of a magnetic poly(methyl methacrylate) core with a poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) shell cross-linked with N, N'-methylenebisacrylamide were prepared by miniemulsion polymerization. The particles were shown to have an average hydrodynamic diameter of 317 nm at 18°C, which decreased to 277 nm at 41°C due to the collapse of the thermo-responsive shell. The particles were superparamagnetic in behavior and exhibited a saturation magnetization of 12.6 emu/g. Subsequently, we evaluated the potential of these negatively charged stimuli-responsive magnetic particles in the purification of a monoclonal antibody from a diafiltered CHO cell culture supernatant by cation exchange. The adsorption of antibodies onto P(NIPAM-co-AA)-coated nanoparticles was highly selective and allowed for the recovery of approximately 94% of the mAb. Different elution strategies were employed providing highly pure mAb fractions with host cell protein (HCP) removal greater than 98%. By exploring the stimuli-responsive properties of the particles, shorter magnetic separation times were possible without significant differences in product yield and purity. PMID:23420794

Borlido, Luís; Moura, Leila; Azevedo, Ana M; Roque, Ana C A; Aires-Barros, Maria R; Farinha, José Paulo S

2013-06-01

283

Antigen production for monoclonal antibody generation.  

PubMed

The quality of the target antigen is very important in order to generate a good antibody, in particular when binding to a conformational epitope is desired. The use of mammalian cells for recombinant protein expression provides an efficient machinery for the correct folding and posttranslational modification of proteins. In this chapter, we describe a process to rapidly generate semi-stable human cell lines secreting a recombinant protein of interest into the culture medium. Simple disposable bioreactors that can be used in any standard cell culture laboratory enable the production of recombinant protein in the multi-milligram range. The protein can be readily purified from the culture supernatant by immobilized metal affinity chromatography. In addition, by inserting a tag recognized by a co-expressed biotin ligase, the protein can be biotinylated during the secretion process. This greatly facilitates the immobilization of the protein for assay development or for antibody isolation using in vitro selection technologies. PMID:24515456

Magistrelli, Giovanni; Malinge, Pauline

2014-01-01

284

Targeted siRNA Delivery and mRNA Knockdown Mediated by Bispecific Digoxigenin-binding Antibodies.  

PubMed

Bispecific antibodies (bsAbs) that bind to cell surface antigens and to digoxigenin (Dig) were used for targeted small interfering RNA (siRNA) delivery. They are derivatives of immunoglobulins G (IgGs) that bind tumor antigens, such as Her2, IGF1-R, CD22, and LeY, with stabilized Dig-binding variable domains fused to the C-terminal ends of the heavy chains. siRNA that was digoxigeninylated at its 3'end was bound in a 2:1 ratio to the bsAbs. These bsAb-siRNA complexes delivered siRNAs specifically to cells that express the corresponding antigen as demonstrated by flow cytometry and confocal microscopy. The complexes internalized into endosomes and Dig-siRNAs separated from bsAbs, but Dig-siRNA was not released into the cytoplasm; bsAb-targeting alone was thus not sufficient for effective mRNA knockdown. This limitation was overcome by formulating the Dig-siRNA into nanoparticles consisting of dynamic polyconjugates (DPCs) or into lipid-based nanoparticles (LNPs). The resulting complexes enabled bsAb-targeted siRNA-specific messenger RNA (mRNA) knockdown with IC(50) siRNA values in the low nanomolar range for a variety of bsAbs, siRNAs, and target cells. Furthermore, pilot studies in mice bearing tumor xenografts indicated mRNA knockdown in endothelial cells following systemic co-administration of bsAbs and siRNA formulated in LNPs that were targeted to the tumor vasculature.Molecular Therapy - Nucleic Acids (2012) 1, e45; doi:10.1038/mtna.2012.39; published online 18 September 2012. PMID:23344238

Schneider, Britta; Grote, Michael; John, Matthias; Haas, Alexander; Bramlage, Birgit; Ickenstein, Ludger M; Jahn-Hofmann, Kerstin; Bauss, Frieder; Cheng, Weijun; Croasdale, Rebecca; Daub, Karin; Dill, Simone; Hoffmann, Eike; Lau, Wilma; Burtscher, Helmut; Ludtke, James L; Metz, Silke; Mundigl, Olaf; Neal, Zane C; Scheuer, Werner; Stracke, Jan; Herweijer, Hans; Brinkmann, Ulrjch

2012-01-01

285

Delivery of the ribosome-inactivating protein, gelonin, to lymphoma cells via CD22 and CD38 using bispecific antibodies.  

PubMed Central

It is well established that bispecific antibodies (BsAbs) can be used effectively in targeting the ribosome-inactivating protein (RIP), saporin, against neoplastic B cells. We have now extended this delivery system for use with gelonin. By measuring antigen-binding characteristics and epitope mapping a panel of anti-gelonin MAbs using the IAsys resonant mirror bisensor, we were able to rapidly select the most suitable for making BaAbs. The Fab' fragments from these MAbs were chemically conjugated with Fab' from either anti-CD22 or anti-CD38. Cytotoxicity assays showed that BsAbs were highly efficient at delivering gelonin to cultured Daudi cells and achieved levels of toxicity which correlated closely with the affinity of the BsAbs. Using pairs of anti-CD22 BsAbs we were able to generate bivalent BsAb-gelonin complexes which achieved IC50 values of 2 x 10(-11) M gelonin, a potency which is equivalent to that reached by saporin in this targeting system. However, because gelonin is 5-10 times less toxic than saporin, the therapeutic ratio for gelonin is superior, making it potentially a more useful agent for human treatment. Cytotoxicity assays and kinetic analysis showed that targeting gelonin via CD38 was 2-5 times less effective than delivery through CD22. However, with a pair of BsAbs designed to co-target gelonin via CD22 and CD38, the cytotoxicity achieved equalled that obtained with a pair of anti-CD22 BsAbs (IC50 = 1 x 10(-11) M). This important result suggests that the anti-CD38 helps bind the gelonin to the cell and is then 'dragged' or 'piggy-backed' into the cell by the anti-CD22 BsAb. The implication of these findings for cancer therapy is discussed.

French, R. R.; Penney, C. A.; Browning, A. C.; Stirpe, F.; George, A. J.; Glennie, M. J.

1995-01-01

286

Anti-CD3 x Anti-GD2 Bispecific Antibody Redirects T-Cell Cytolytic Activity to Neuroblastoma Targets  

PubMed Central

Background The ganglioside GD2 is an attractive target for immunotherapy of neuroectodermal tumors. We tested a unique bispecific antibody anti-CD3 × anti-GD2 (3F8BiAb) for its ability to redirect activated T cells (ATC) to target GD2-positive neuroblastomas. Procedure ATC were generated from normal human peripheral blood mononuclear cells (PBMC) by stimulating the PBMC with OKT3 and expanding the T cells in the presence of interleukin 2 (IL-2) for 14 days. ATC were armed with 3F8BiAb (100 ng/106 cells) or Her2BiAb (50 ng/106 cells) prior to use. 3F8 BiAb were tested for its dual-binding specificity to GD2 expressed on cancer cell lines and CD3 expressed on ATC. 3F8BiAb-armed ATC were further tested ex vivo for their cytotoxicity against GD2 positive tumor targets and its ability to induce cytokine response upon binding to targets. Results GD2 expression in neuroblastoma cells was confirmed by FACS analysis. Specific binding of 3F8BiAb to the tumor targets as well as to ATC was confirmed by FACS analysis. 3F8BiAb-armed ATC exhibited specific killing of GD2 positive neuroblastoma cell lines significantly above unarmed ATC (P < 0.001). GD2BiAb-armed ATC secreted significantly higher levels of Th1 cytokines and chemokines compared to unarmed ATC (P < 0.001). Conclusions These preclinical findings support the potential of a novel immunotherapeutic approach to target T cells to neuroblastoma.

Yankelevich, Maxim; Kondadasula, Sri Vidya; Thakur, Archana; Buck, Steven; Cheung, Nai-Kong V.; Lum, Lawrence G.

2013-01-01

287

Targeting cytomegalovirus-infected cells using T cells armed with anti-CD3 × anti-CMV bispecific antibody.  

PubMed

Human cytomegalovirus (CMV) reactivation and infection can lead to poor outcomes after allogeneic stem cell transplantation. We hypothesized that anti-CD3 activated T cells (ATCs) armed with chemically heteroconjugated anti-CD3 × polyclonal anti-CMV bispecific antibody (CMVBi) will target and eliminate CMV-infected cells. Arming doses of CMVBi as low as 0.01 ng/10(6) ATCs was able to mediate specific cytotoxicity (SC) directed at CMV-infected target cells significant above unarmed ATCs at mutiplicities of infection (MOI) between 0.01 and 1. At effector-to-target ratios (E:T) of 25:1, 12.5:1, 6.25:1, and 3.125:1, armed ATCs significantly enhanced killing of CMV-infected targets compared with unarmed ATCs. At an MOI of 1.0, the mean % SC directed at CMV-infected targets cells for CMVBi-armed ATCs at E:T of 3.12, 6.25, and 12.5 were 79%, 81%, and 82%, respectively; whereas the mean % SC for unarmed ATCs at the same E:T were all <20%. ATCs, Cytogam(®), or CMVBi alone did not lyse uninfected or CMV-infected targets. Co-cultures of CMVBi-armed ATCs with CMV-infected targets induced cytokine and chemokine release from armed ATCs. This nonmajor histocompatibility complex restricted strategy for targeting CMV could be used to prevent or treat CMV infections after allogeneic stem cell transplantation or organ transplantation. PMID:22313635

Lum, Lawrence G; Ramesh, Mayur; Thakur, Archana; Mitra, Subhashis; Deol, Abhinav; Uberti, Joseph P; Pellett, Philip E

2012-07-01

288

Targeted siRNA Delivery and mRNA Knockdown Mediated by Bispecific Digoxigenin-binding Antibodies  

PubMed Central

Bispecific antibodies (bsAbs) that bind to cell surface antigens and to digoxigenin (Dig) were used for targeted small interfering RNA (siRNA) delivery. They are derivatives of immunoglobulins G (IgGs) that bind tumor antigens, such as Her2, IGF1-R, CD22, and LeY, with stabilized Dig-binding variable domains fused to the C-terminal ends of the heavy chains. siRNA that was digoxigeninylated at its 3?end was bound in a 2:1 ratio to the bsAbs. These bsAb–siRNA complexes delivered siRNAs specifically to cells that express the corresponding antigen as demonstrated by flow cytometry and confocal microscopy. The complexes internalized into endosomes and Dig-siRNAs separated from bsAbs, but Dig-siRNA was not released into the cytoplasm; bsAb-targeting alone was thus not sufficient for effective mRNA knockdown. This limitation was overcome by formulating the Dig-siRNA into nanoparticles consisting of dynamic polyconjugates (DPCs) or into lipid-based nanoparticles (LNPs). The resulting complexes enabled bsAb-targeted siRNA-specific messenger RNA (mRNA) knockdown with IC50 siRNA values in the low nanomolar range for a variety of bsAbs, siRNAs, and target cells. Furthermore, pilot studies in mice bearing tumor xenografts indicated mRNA knockdown in endothelial cells following systemic co-administration of bsAbs and siRNA formulated in LNPs that were targeted to the tumor vasculature.

Schneider, Britta; Grote, Michael; John, Matthias; Haas, Alexander; Bramlage, Birgit; lckenstein, Ludger M; Jahn-Hofmann, Kerstin; Bauss, Frieder; Cheng, Weijun; Croasdale, Rebecca; Daub, Karin; Dill, Simone; Hoffmann, Eike; Lau, Wilma; Burtscher, Helmut; Ludtke, James L; Metz, Silke; Mundigl, Olaf; Neal, Zane C; Scheuer, Werner; Stracke, Jan; Herweijer, Hans; Brinkmann, Ulrjch

2012-01-01

289

Immunohistochemical identification of cytotoxic lymphocytes using human perforin monoclonal antibody.  

PubMed Central

Perforin is a potent cytolytic pore-forming protein expressed in cytoplasmic granules of cytotoxic T lymphocytes and natural killer cells. A new monoclonal antibody raised against human perforin was used to detect both in vitro and in vivo perforin expression in cytotoxic cells. Immunohistochemical analysis of human peripheral blood mononuclear cells cultured in recombinant interleukin-2 (rIL-2) showed strong granular cytoplasmic staining of the IL-2 activated cytotoxic cells. Fresh-frozen tissue sections from patients with heart allograft rejection were also stained. Strong granular cytoplasmic staining of the mononuclear inflammatory infiltrate characteristic for perforin in cardiac allograft rejection was observed. The detection and quantitative analysis of perforin-associated cytotoxic cells by the human anti-perforin monoclonal antibody will help to evaluate the significance of these functionally distinct cytotoxic cells in human tissue. Images Figure 1

Hameed, A.; Olsen, K. J.; Cheng, L.; Fox, W. M.; Hruban, R. H.; Podack, E. R.

1992-01-01

290

Production and Characterization of Monoclonal Antibodies against Aflatoxin B1.  

PubMed

In this article, we embarked on production of mouse monoclonal antibodies against aflatoxin B1 which is the most commonly occurring fungal toxin in food and feed products. After immunization and fusion with myloma cells, two stable clones (A218 and B319) were selected. Isotyping showed that these monoclonal antibodies (mAbs) were IgG2b with kappa light chains. The affinity of A218 and B319 clons were 5×10(11) M(-1) and 6×10(9) M(-1), respectively. Competitive indirect ELISA results indicated these mAbs had complete (100%) cross-reaction with four major types of aflatoxins: B1, B2, G1, and G2. These mAbs could be used for immunoassay measurement of aflatoxins with high affinity and low detection limits. PMID:24350626

Soukhtanloo, Mohammad; Talebian, Elham; Golchin, Mehdi; Mohammadi, Mojgan; Amirheidari, Bagher

2014-01-01

291

Desensitizations for chemotherapy and monoclonal antibodies: indications and outcomes.  

PubMed

Acute infusion reactions to both chemotherapeutic agents and humanized monoclonal antibodies can occur, which may limit therapeutic options for treatment of malignancies and chronic inflammatory diseases. Many of these acute infusion reactions are consistent with a type I hypersensitivity reaction, including anaphylaxis. If a patient experiences a significant acute infusion reaction, often the recommendation is to discontinue the medication and find an alternative agent. However, the "second-line" agent may be more toxic or inferior. If the reaction is likely a type I or type IV hypersensitivity reaction, one option is to undergo desensitization to the offending drug. Drug desensitization is the process of readministering a needed drug in incremental doses over hours or days until a full therapeutic dose is tolerated. This article will review the current literature on indications and outcomes for drug desensitization in the management of allergy to either chemotherapeutic agents or monoclonal antibodies. PMID:24994467

Hsu Blatman, Karen S; Castells, Mariana C

2014-08-01

292

A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities.  

PubMed Central

We report a novel approach to the generation of monoclonal antibodies based on the molecular cloning and expression of immunoglobulin variable region cDNAs generated from single rabbit or murine lymphocytes that were selected for the production of specific antibodies. Single cells secreting antibodies for a specific peptide either from gp116 of the human cytomegalovirus or from gp120 of HIV-1 or for sheep red blood cells were selected using antigen-specific hemolytic plaque assays. Sheep red blood cells were coated with specific peptides in a procedure applicable to any antigen that can be biotinylated. Heavy- and light-chain variable region cDNAs were rescued from single cells by reverse transcription-PCR and expressed in the context of human immunoglobulin constant regions. These chimeric murine and rabbit monoclonal antibodies replicated the target specificities of the original antibody-forming cells. The selected lymphocyte antibody method exploits the in vivo mechanisms that generate high-affinity antibodies. This method can use lymphocytes from peripheral blood, can exploit a variety of procedures that identify individual lymphocytes producing a particular antibody, and is applicable to the generation of monoclonal antibodies from many species, including humans. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4

Babcook, J S; Leslie, K B; Olsen, O A; Salmon, R A; Schrader, J W

1996-01-01

293

Immunomodulation of transplant rejection using monoclonal antibodies and soluble receptors  

Microsoft Academic Search

The main objective of our studies has been to optimize the effects of monoclonal antibodies (MAbs) and other immunosuppressive reagents to enhance organ graft survival. One such agent is OKT3, a MAb that is directed against the CD3 component of the human T-cell receptor (TCR) complex. Treatment of a rejection episode with OKT3 results in a rapid and efficient clearing

Maria-Luisa Alegre; Deborah J. Lenschow; Jeffrey A. Bluestone

1995-01-01

294

Reviews: Interspecies Scaling of Therapeutic Monoclonal Antibodies: Initial Look  

Microsoft Academic Search

The authors evaluated interspecies scaling for the prediction of human clearance of 18 therapeutic monoclonal antibodies (mAbs). Human and monkey\\/chimpanzee data of 14 mAbs were classified based on the targeted antigens (soluble or membrane bound). Simple allometry and\\/or a time-invariant method (elementary Dedrick plot) were performed. Results indicate that human clearance might be accurately predicted from monkey data for mAbs

Jie Ling; Honghui Zhou; Qun Jiao; Hugh M. Davis

2009-01-01

295

Monoclonal antibodies recognizing single amino acid substitutions in hemoglobin  

Microsoft Academic Search

Four monoclonal antibodies (mAb) to non-human primate hemoglobin referred to as Cap-4, Cap-5, Rh-2, and Rh-4, and two mAb to human hemoglobin, referred to as H-1 and H-3 were isolated and were partially characterized. Binding studies with these mAb on a panel of hemoglobins and isolated ..cap alpha.. and ..beta.. globin chains revealed a unique reactivity pattern for each mAb.

L. H. Stanker; E. Branscomb; M. Vanderlaan; R. H. Jensen

1986-01-01

296

Neutralizing monoclonal antibodies directed to infectious bovine rhinotracheitis virus  

Microsoft Academic Search

Summary Infectious bovine rhinotracheitis virus (IBRV) has been shown in this report to have thirty-three polypeptides. Ten of the eleven polypeptides which can be labeled with (3H)-glucosamine are located on the surface of the virus since they can be surface labeled with sodium boro(3H)hydride. In order to define the immunologically important viral proteins, monoclonal antibodies were prepared against the virus

L. W. S. Chang; Y. C. Zee; R. F. Pritchett; A. A. Ardans

1986-01-01

297

Monoclonal Antibodies in the Treatment of Malignant Lymphomas  

Microsoft Academic Search

Non-Hodgkin's lymphoma is a heterogeneous group of B- and T-cell cancers, with a large variety of patterns of growth, clinical\\u000a presentations, and responses to treatment. Outcome depends on histological subtype, tumor characteristics, host responses,\\u000a and treatment. About 90 percent of lymphomas have a B-cell phenotype and for them recent therapeutic progress came from the\\u000a introduction of monoclonal antibodies (MAb) alone

Bertrand Coiffier

298

Analysis of thermal stability of soya globulins using monoclonal antibodies  

Microsoft Academic Search

The epitopes of two monoclonal antibodies (Mabs), one raised to soya glycinin (IFRN 0025) and one to ?-conglycinin (IFRN 0089), have been defined. The epitope of 0025 corresponds to residues 86–104 of the acidic polypeptide of glycinin A1aB1b and lies at the C terminus of the proteolytic intermediate known as glycinin-T, whilst that of 0089 corresponds to residues 78–84 in

Ling Huang; E. N. Clare Mills; Jane M. Carter; Michael R. A. Morgan

1998-01-01

299

Radiolabeled monoclonal antibodies: radiochemical pharmacokinetic and clinical challenges  

SciTech Connect

Four basic science articles and two special contributions included in this issue of the journal address some of the persistent challenges that continue to moderate the progress of radioimmunoimaging and therapy. This editorial will review, and expand upon, several points brought out in these experiments. One focus will be on the chemistry issues, thereby supplementing the excellent review article on monoclonal antibodies presented by Keenan, as well as some discussion about clinical considerations.

Gobuty, A.H.; Kim, E.E.; Weiner, R.E.

1985-05-01

300

Trypanosoma cruzi: amastigote polymorphism defined by monoclonal antibodies  

Microsoft Academic Search

We have raised monoclonal antibodies (mAbs) directed towards amastigote forms of Trypanosoma cruzi, and shown that mAbs 1D9 and 4B9 are carbohydrate while mAb 4B5 activity is resistant to periodate oxidation of the antigen. Here we used an ELISA to quanti- tate and compare the expression of surface epitopes on fixed parasites among different parasite isolates. The expression of markers

N. V. Verbisck; S. Da-Silva; R. A. Mortara

1998-01-01

301

Monoclonal Antibody Therapy for B-Cell Lymphoma  

Microsoft Academic Search

Treatment strategies and outcomes for patients with non-Hodgkin’s lymphoma (NHL) are undergoing rapid and important evolution.\\u000a We have recently witnessed the advent of novel targeted therapies, such as gene therapies, active immunotherapies, antisense\\u000a therapies, new small molecules and biologicals, and monoclonal antibodies (MoAbs). The first MoAb approved for the treatment\\u000a of cancer, rituximab, was approved in 1997 and has been

Antonio J. Grillo-López

2002-01-01

302

Production and characterization of monoclonal antibodies to a grouper iridovirus  

Microsoft Academic Search

A panel of six monoclonal antibodies (mAbs) against a grouper iridovirus (SGIV) were produced by immunization of Balb\\/c mice with purified virus preparations. Isotyping test revealed all the mAbs were IgG1. None of the mAbs possessed the ability to neutralize SGIV in cell cultures. Western blot showed that 4 mAbs reacted with 2 SGIV proteins at molecular mass of approximately

Chengyin Shi; Qi Wei; Karina Yew Hoong Gin; Toong Jin Lam

2003-01-01

303

Production of monoclonal antibodies against quail Pax-1.  

PubMed

Monoclonal antibodies (MAbs) against quail Pax-1 were generated using a fusion protein of the C-terminal part of Pax-1 and glutathione S-transferase. The MAbs generated could detect quail Pax-1 protein by Western blotting and immunoprecipitate it from whole cell extracts of quail embryos, indicating their usefulness for biochemical analyses of Pax-1 protein function in development or oncogenesis. PMID:9064290

Kaneko, K; Taniguchi, M; Isono, K; Koseki, H

1996-02-01

304

Alzheimer paired helical filaments: Identification of polypeptides with monoclonal antibodies  

Microsoft Academic Search

Paired helical filaments (PHF) were isolated from autopsied brain of cases of Alzheimer dementia, and their polypeptides were identified with monoclonal antibodies to PHF by Western blots. The PHF polypeptide profile consisted of several bands with a size difference of less than 5 kilodalton (kDa) between adjacent bands; the most prominent bands were in the 45–62 kDa region. These PHF

I. Grundke-Iqbal; G. P. Wang; K. Iqbal; Y.-C. Tung; H. M. Wisniewski

1985-01-01

305

Monoclonal antibodies directed against surface molecules of multicell spheroids  

NASA Technical Reports Server (NTRS)

The objective of this project is to generate a library of monoclonal antibodies (MAbs) to surface molecules of mammalian tumor and transformed cells grown as multicell spheroids (MCS). These MCS are highly organized, three dimensional multicellular structures which exhibit many characteristics of in vivo organized tissues not found in conventional monolayer or suspension culture; therefore, MCS make better in vitro model systems to study the interactions of mammalian cells. Additionally, they provide a functional assay for surface adhesion molecules.

Martinez, Andrew O.

1993-01-01

306

Monoclonal antibodies as a tool for phylogenetic studies of major histocompatibility antigens and ? 2 -microglobulin  

Microsoft Academic Search

The cross-reactivity of several monoclonal antibodies recognizing monomorphic determinants of human HLA-A, B, C, and DR antigens and human ß2-microglobulin (ß2m) has been studied on peripheral blood leukocytes in 24 different species. An monoclonal HLA-A-, B-, and C-specific antibody and four monoclonal HLA-DR-specific antibodies cross-reacted with cells from all the primate species tested. Furthermore, antibodies HLA-DR-specific were positive with peripheral

Jean-Luc Teillaud; Denis Crevat; Patrick Chardon; Jorge Kalil; Cécile Goujet-Zalc; Guy Mahouy; Marcel Vaiman; Marc Fellous; Donald Piou

1982-01-01

307

I. Induction of proteinuria in the rat by a monoclonal antibody against SGP115\\/107  

Microsoft Academic Search

I. Induction of proteinuria in the rat by a monoclonal antibody against SGP-115\\/107. The role of individual antigenic determinants in the pathogenesis of antibody-mediated glomerular injury is, at present, incompletely understood. This study was designed to compare the effect of monoclonal antibodies upon binding to three distinct antigenic determinants present in the glomerular capillary wall. Ten monoclonal antibodies were tested

Donna L Mendrick; Helmut G Rennke

1988-01-01

308

[Targeted therapies including monoclonal antibodies for connective tissue diseases].  

PubMed

Recent advance of targeted therapies including monoclonal antibodies and fusion proteins has allowed effective strategies in the treatment of rheumatoid arthritis. And now, TNF inhibitors are broadly used for rheumatoid arthritis and prevent the disease progression. Meanwhile, B cell targeted therapies and anti-interleukin-6 receptor antibody treatment are not only used for second line biological agents for rheumatoid arthritis, but also expected for the treatments of various autoimmune diseases. Recent year, some of novel small molecules, which inhibit the signal transduction of various surface receptors of immune cells, are in clinical trials. These drugs will be a breakthrough for the treatment of some autoimmune disorders. PMID:19280937

Hayashi, Taichi; Sumida, Takayuki

2009-03-01

309

Production of Bartonella Genus-Specific Monoclonal Antibodies  

PubMed Central

Monoclonal antibodies (MAbs) which react with heat-resistant proteins with molecular masses of 32 to 33 kDa of 14 different Bartonella species were produced. These antibodies did not react with antigens of 26 diverse bacterial strains by microimmunofluorescence assay except MAb B3D4, which reacted with Chlamydia psittaci and Chlamydia trachomatis at low titers. The identification of a common Bartonella antigenic protein will make it possible to later produce a diagnostic antigen by cloning and expressing it in Escherichia coli. Moreover, these MAbs allow all Bartonella species to be identified to the genus level.

Liang, Zhongxing; La Scola, Bernard; Lepidi, Hubert; Raoult, Didier

2001-01-01

310

Screening individual hybridomas by microengraving to discover monoclonal antibodies  

PubMed Central

The demand for monoclonal antibodies (mAbs) in biomedical research is significant, but the current methodologies used to discover them are both lengthy and costly. Consequently, the diversity of antibodies available for any particular antigen remains limited. Microengraving is a soft lithographic technique that provides a rapid and efficient alternative for discovering new mAbs. This protocol describes how to use microengraving to screen mouse hybridomas to establish new cell lines producing unique mAbs. Single cells from a polyclonal population are isolated into an array of microscale wells (~105 cells per screen). The array is then used to print a protein microarray, where each element contains the antibodies captured from individual wells. The antibodies on the microarray are screened with antigens of interest, and mapped to the corresponding cells, which are then recovered from their microwells by micromanipulation. Screening and retrieval require approximately 1–3 d (9–12 d including the steps for preparing arrays of microwells).

Ogunniyi, Adebola O; Story, Craig M; Papa, Eliseo; Guillen, Eduardo; Love, J Christopher

2014-01-01

311

Monoclonal antibodies to human vitamin D-binding protein.  

PubMed Central

Monoclonal antibodies to vitamin D-binding protein isolated from human serum have been produced. The antibodies obtained have been shown to be specific for human vitamin D-binding protein by three independent assays. The antibodies recognize human vitamin D-binding protein specifically in an enzyme-linked immunosorbent assay. Human vitamin D-binding protein is detected specifically in both pure and crude samples by a radiometric immunosorbent assay (RISA) and by an immunoprecipitation assay. The anti-human vitamin D-binding protein antibodies cross-react with monkey and pig vitamin D-binding protein, but not with vitamin D-binding protein from rat, mouse, or chicken, as determined by the RISA and immunoprecipitation assays. Images

Pierce, E A; Dame, M C; Bouillon, R; Van Baelen, H; DeLuca, H F

1985-01-01

312

Monoclonal antibodies in the treatment of pancreatic cancer.  

PubMed

Human pancreatic cancer is a malignant disease with almost equal incidence and mortality. Effective diagnostic and therapeutic strategies are still urgently needed to improve its survival rate. With advances in structural and functional genomics, recent work has focused on targeted molecular therapy using monoclonal antibodies. This review summarizes the target molecules on the tumor cell surface and normal tissue stroma, which are related to pancreatic cancer oncogenesis, tumor growth or resistance to chemotherapy, as well as molecules involved in regulating inflammation and host immunoresponses. Targeted molecules include cell-surface receptors, such as the EGF receptor, HER2, death receptor 5 and IGF-1 receptor. Effects of monoclonal antibodies against these target molecules alone or in combination with chemotherapy, small-molecule signal transduction inhibitors, or radiation therapy are also discussed. Also discussed are the use of toxin or radioisotope conjugates, and information relating to the use of these targeting agents in pancreatic cancer clinical trials. Although targeted molecular therapy with monoclonal antibodies has made some progress in pancreatic cancer treatment, especially in preclinical studies, its clinical application to improve the survival rate of pancreatic cancer patients requires further investigation. PMID:20046965

Huang, Zhi-Qiang; Buchsbaum, Donald J

2009-03-01

313

[Identification of L. donovani promastigotes from Phlebotomus by monoclonal antibody].  

PubMed

It has been reported by the authors that monoclonal antibody L12G9 produced from target antigens of L. donovani promastigotes, was very useful for detecting promastigotes from artificially infected sandflies. In the present study, detection of promastigotes from artificially infected sandflies by McAb showed that the positive rate correlated with the infection duration of sandflies. 4 days after feeding on infected Chinese hamsters, the sandflies were lightly infected with L. donovani promastigotes with a positive rate of 15.9%, but 10 days later, the sandflies were heavily infected, the positive rate being 100%. Observation has been made on the relationship between the number of promastigotes and mouse blood dilution. The results showed that satisfactory results could be obtained by using monoclonal antibodies in the detection of L. donovani, the number of promastigotes should be over 1 x 10(7)/ml, and the blood meals in sandflies completely digested. If very few promastigotes were present in naturally infected sandflies before identification by monoclonal antibody, the parasites must be grown in NNN medium. Positive result could then be obtained. PMID:2099257

Qu, J Q; Xu, Y X; Guan, L R; Bao, Y F

1990-01-01

314

Monoclonal antibodies in the treatment of pancreatic cancer  

PubMed Central

Human pancreatic cancer is a malignant disease with almost equal incidence and mortality. Effective diagnostic and therapeutic strategies are still urgently needed to improve its survival rate. With advances in structural and functional genomics, recent work has focused on targeted molecular therapy using monoclonal antibodies. This review summarizes the target molecules on the tumor cell surface and normal tissue stroma, which are related to pancreatic cancer oncogenesis, tumor growth or resistance to chemotherapy, as well as molecules involved in regulating inflammation and host immunoresponses. Targeted molecules include cell-surface receptors, such as the EGF receptor, HER2, death receptor 5 and IGF-1 receptor. Effects of monoclonal antibodies against these target molecules alone or in combination with chemotherapy, small-molecule signal transduction inhibitors, or radiation therapy are also discussed. Also discussed are the use of toxin or radioisotope conjugates, and information relating to the use of these targeting agents in pancreatic cancer clinical trials. Although targeted molecular therapy with monoclonal antibodies has made some progress in pancreatic cancer treatment, especially in preclinical studies, its clinical application to improve the survival rate of pancreatic cancer patients requires further investigation.

Huang, Zhi-Qiang; Buchsbaum, Donald J

2009-01-01

315

Phase 1 trial of the novel bispecific molecule H22xKi-4 in patients with refractory Hodgkin lymphoma  

Microsoft Academic Search

CD30 is an excellent target for immuno- therapy of Hodgkin lymphoma (HL) be- cause it is overexpressed on Hodgkin and Reed-Sternberg cells. We developed a novel bispecific molecule (BSM) consist- ing of F(ab) fragments derived from the murine anti-CD30 monoclonal antibody (MoAb) Ki-4 and the humanized CD64- specific MoAb H22. In vitro experiments of H22xKi-4 demonstrated specific phago- cytosis of

Peter Borchmann; Roland Schnell; Irene Fuss; Oliver Manzke; Thomas Davis; Lionel D. Lewis; Detlev Behnke; Claudia Wickenhauser; Petra Schiller; Volker Diehl; Andreas Engert

2002-01-01

316

Induction of regular cytolytic T cell synapses by bispecific single-chain antibody constructs on MHC class I-negative tumor cells.  

PubMed

Certain bispecific single-chain antibody constructs exhibit an extraordinary potency for polyclonal T cell engagement and target cell lysis. Here we studied the structural basis for this potency, using laser scanning confocal microscopy. Cytolytic human T cell synapses could be triggered either by addition of a specific peptide antigen or an Ep-CAM-/CD3-bispecific T cell engager (BiTE). Both kinds of synapses showed a comparable distribution of all protein markers investigated. Two other BiTEs constructed from different Ep-CAM-specific antibodies gave similar results. BiTEs could also induce lytic synapses between human T cells and a MHC class I-negative, Ep-CAM cDNA-transfected cell line resulting in potent target cell lysis. This shows that certain T cell recognition molecules on target cells are dispensable for synapse formation and BiTE activity, and suggests that BiTE-activated polyclonal T cells may ignore major immune evasion mechanisms of tumor cells in vivo, such as loss of MHC class I expression. PMID:16360021

Offner, Sonja; Hofmeister, Robert; Romaniuk, Andrea; Kufer, Peter; Baeuerle, Patrick A

2006-02-01

317

Intravesical administration of radiolabeled antitumor monoclonal antibody in bladder carcinoma  

SciTech Connect

Tumor associated AUA1 monoclonal antibody and 11.4.1. nonspecific monoclonal antibody, which does not react with human tissues, were radiolabeled with 111In and administered intravesically to 23 patients undergoing cystoscopy for bladder carcinoma. The antibody solution remained in the bladder for 1 h and then was washed out prior to cystoscopy. Tumor and nontumor samples were obtained during cystoscopy and were counted in a gamma counter. Conventional and immunoperoxidase staining with both antibodies were also performed. AUA1 reacted with all bladder carcinomas while 11.4.1. was negative in all cases. The mean uptake of AUA1 at 2, 24, and 48 h after the instillation (expressed as 10(3) x percentage of injected dose/g of tissue) was: 6.12 +/- 5.50 (SD), 1.70 +/- 2.57, 0.30 +/- 0.17 in the tumors and 0.32 +/- 0.50, 0.22 +/- 0.30, 0 in the nontumor areas, and for 11.4.1. it was: 0.075 +/- 0.075, 0.025 +/- 0.025 in the tumors and 0.30 +/- 0.42, 0.15 +/- 0.26 in the nontumor areas. The uptake of AUA1 by the tumors correlated with the tumor grade. There was no radioactivity in the blood at 2 h, and at 1, 2, and 3 days after the instillation. Our results indicate that intravesical administration of radiolabeled monoclonal antibody AUA1 targets selectively to tumor tissue without any significant normal tissue uptake. This finding might allow the development of a nontoxic and specific therapeutic approach for superficial bladder carcinoma.

Bamias, A.; Keane, P.; Krausz, T.; Williams, G.; Epenetos, A.A. (Imperial Cancer Research Fund, London (England))

1991-01-15

318

Influence of unlabeled monoclonal anti-mouse antibody on the clearance rate of radiolabeled mouse monoclonal antibody  

Microsoft Academic Search

High blood background levels of intact radiolabeled monoclonal antibody (MoAb) after intravenous (iv) injection are problematic. The injection of unlabeled polyclonal antimouse Abs following injection with labeled MoAbs produces accelerated MoAb clearance. This study evaluates a Mo antimouse Ab for efficacy of accelerating radio MoAb clearance. HB-58 is a rat\\/mouse MoAb which binds strongly to mouse kappa light chains present

R. L. Wahl; L. Laino; G. Jackson; S. Fisher; W. H. Beierwaltes

1985-01-01

319

Mouse Monoclonal Antibodies to Anthrax Edema Factor Protect against Infection ?  

PubMed Central

Bacillus anthracis is the causative agent of anthrax, and the tripartite anthrax toxin is an essential element of its pathogenesis. Edema factor (EF), a potent adenylyl cyclase, is one of the toxin components. In this work, anti-EF monoclonal antibodies (MAb) were produced following immunization of mice, and four of the antibodies were fully characterized. MAb 3F2 has an affinity of 388 pM, was most effective for EF detection, and appears to be the first antibody reported to neutralize EF by binding to the catalytic CB domain. MAb 7F10 shows potent neutralization of edema toxin activity in vitro and in vivo; it targets the N-terminal protective antigen binding domain. The four MAb react with three different domains of edema factor, and all were able to detect purified edema factor in Western blot analysis. None of the four MAb cross-reacted with the lethal factor toxin component. Three of the four MAb protected mice in both a systemic edema toxin challenge model and a subcutaneous spore-induced foreleg edema model. A combination of three of the MAb also significantly delayed the time to death in a third subcutaneous spore challenge model. This appears to be the first direct evidence that monoclonal antibody-mediated neutralization of EF alone is sufficient to delay anthrax disease progression.

Leysath, Clinton E.; Chen, Kuang-Hua; Moayeri, Mahtab; Crown, Devorah; Fattah, Rasem; Chen, Zhaochun; Das, Suman R.; Purcell, Robert H.; Leppla, Stephen H.

2011-01-01

320

Detection of chromogranin in neuroendocrine cells with a monoclonal antibody.  

PubMed Central

A monoclonal antibody ( LK2H10 ) produced against a human pheochromocytoma reacted immunohistochemically with 126 normal and neoplastic endocrine tissues with secretory granules which were formalin-fixed and paraffin-embedded. Antibody LK2H10 did not react with 46 other endocrine tissues or tumors without secretory granules nor with 113 normal and neoplastic nonendocrine cells and tumors. Tumors with abundant secretory granules showed intense and diffuse staining, and tumors with few granules, such as Merkel cell carcinomas, neuroblastomas, and small cell carcinomas of lung, showed focal staining. Antibody LK2H10 did not react with melanomas, nevi, posterior pituitary, peripheral nerve tissues, or neurons. The target structure of LK2H10 was identified as human chromogranin, of which the major fraction was chromogranin A (mol wt 68,000 daltons). Preabsorption with purified chromogranin A blocked immunoperoxidase staining by LK2H10 in normal adrenal medulla, in the anterior pituitary, and in a pheochromocytoma. Ultrastructural immunohistochemistry with LK2H10 showed that chromogranin was present in cytoplasmic secretory granules. These results indicate that chromogranin is widely distributed in the secretory granules of most polypeptide-producing endocrine tissues, and it is readily detected with the use of monoclonal antibody LK2H10 . The detection of this marker can be very helpful as a diagnostic aid for neuroendocrine cells and tumors. Images Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 1 Figure 2 Figure 3 Figure 10 Figure 11

Wilson, B. S.; Lloyd, R. V.

1984-01-01

321

Monoclonal antibodies for detecting bone marrow invasion by neuroblastoma.  

PubMed Central

One hundred and sixty six bone marrow aspirates from children with abdominal neuroblastoma were independently examined for the presence of neuroblastoma cells by conventional haematological staining techniques and by indirect immunofluorescence using one or more of five monoclonal antibodies. Results using the two methods were evaluated by comparison with the results of bone marrow, bone and lymph node biopsy specimens taken at the same time, and in the light of the child's clinical course. One antibody, UJ13A, was found to have 98.5% specificity and 85.7% sensitivity, compared with sensitivities for conventional staining of 50% for aspirate alone and of aspirate or trephine biopsy specimen of 71.4%. The remaining antibodies UJ127/11, UJ223/8, UJ181/4 and UJ167/11, with specificities in the range 94-99%, were found to have sensitivities in the range 47-61%, lower than the sensitivity of conventionally stained aspirate or trephine biopsy specimen. Routine immunofluorescence staining with monoclonal antibody UJ13A should increase the sensitivity of detection of metastatic neuroblastoma.

Rogers, D W; Treleaven, J G; Kemshead, J T; Pritchard, J

1989-01-01

322

Phenotypic characterization of Ewing sarcoma cell lines with monoclonal antibodies.  

PubMed

The histogenesis of Ewing sarcoma, the second most frequent bone tumor in humans, remains controversial. Four Ewing cell lines were analyzed by immunological methods. A panel of antibodies directed to T, B, and myelomonocytic markers gave negative results. Surface antigens recognized on Ewing cells were found to be related to the neuroectoderm lineage. Ganglioside GD2, a marker of neuroectodermal tissues and tumors, was present on all lines. These were also stained by the mouse monoclonal antibody HNK-1, which detects a carbohydrate epitope present on several glycoconjugates of the nervous system, including two glycoproteins, the myelin-associated glycoprotein and the neural cell-adhesion molecule (N-CAM), and an acidic glycolipid of the peripheral nervous system. The P61 monoclonal antibody, which reacts with a peptide moiety of N-CAM, and a rabbit antiserum, raised to purified mouse N-CAM and not recognizing the HNK-1-defined epitope, were also reactive. By contrast, all antibodies specific for hematopoietic cell surface antigens were totally negative. Besides these antigenic features, Ewing sarcoma cells are characterized by a specific t(11;22)(q24;q12) translocation also observed in neuroepithelioma, a neuroectodermal tumor, suggesting a possible evolutionary related origin. The recent finding that the human N-CAM gene is located at the vicinity of the breakpoint on chromosome 11 indicates that it might be involved in genetic rearrangements occurring in this region. PMID:3760036

Lipinski, M; Braham, K; Philip, I; Wiels, J; Philip, T; Dellagi, K; Goridis, C; Lenoir, G M; Tursz, T

1986-01-01

323

Modulation of desmin intermediate filament assembly by a monoclonal antibody.  

PubMed

We have used a monoclonal antibody against desmin to examine the assembly of intermediate filaments (IF) from their building blocks, the tetrameric protofilaments. The antibody, designated D76, does not cross react with any other IF proteins (Danto, S.I., and D.A. Fischman. 1984. J. Cell Biol. 98:2179-2191). It binds to a region amino-terminal to cys-324 of avian desmin that is resistant to chymotrypsin and trypsin digestion, and in the electron microscope appears to bind to the ends of tetrameric protofilaments. In combination, these findings suggest that the epitope of the antibody resides at the amino-terminal end of the alpha-helical rod domain. Preincubation of desmin protofilaments with an excess of D76 antibodies blocks their subsequent assembly into IF. In the presence of sub-stoichiometric amounts of antibodies, IF are assembled from protofilaments but they are morphologically aberrant in that (a) they are capped by IgG molecules at one or both ends; (b) they are unraveled to varying degree, revealing a characteristic right-handed helical arrangement of sub-filamentous strands of different diameters. The antibody binds only to the ends but not along the length of desmin IF. The most straightforward explanation for this is that the epitope resides in a part of the desmin molecule that becomes buried within the core of the filament upon polymerization and is therefore inaccessible to the antibody. PMID:2450097

Ip, W

1988-03-01

324

Modulation of desmin intermediate filament assembly by a monoclonal antibody  

PubMed Central

We have used a monoclonal antibody against desmin to examine the assembly of intermediate filaments (IF) from their building blocks, the tetrameric protofilaments. The antibody, designated D76, does not cross react with any other IF proteins (Danto, S.I., and D.A. Fischman. 1984. J. Cell Biol. 98:2179-2191). It binds to a region amino-terminal to cys- 324 of avian desmin that is resistant to chymotrypsin and trypsin digestion, and in the electron microscope appears to bind to the ends of tetrameric protofilaments. In combination, these findings suggest that the epitope of the antibody resides at the amino-terminal end of the alpha-helical rod domain. Preincubation of desmin protofilaments with an excess of D76 antibodies blocks their subsequent assembly into IF. In the presence of sub-stoichiometric amounts of antibodies, IF are assembled from protofilaments but they are morphologically aberrant in that (a) they are capped by IgG molecules at one or both ends; (b) they are unraveled to varying degree, revealing a characteristic right- handed helical arrangement of sub-filamentous strands of different diameters. The antibody binds only to the ends but not along the length of desmin IF. The most straightforward explanation for this is that the epitope resides in a part of the desmin molecule that becomes buried within the core of the filament upon polymerization and is therefore inaccessible to the antibody.

1988-01-01

325

Rapid Characterization of Monoclonal Antibodies using the Piezoelectric Immunosensor  

PubMed Central

Monoclonal antibodies with specificity against the Francisella tularensis outer lipopolysaccharide (LPS) membrane were prepared and characterized using the piezoelectric immunosensor with immobilized LPS antigen from F. tularensis. Signals obtained by the immunosensor were compared with ELISA and similar sensitivity was noticed. Signal of negative controls obtained using the biosensor was below 0.5% of the signal obtained for the selected specific antibody clone 4H3B9D3. Testing of cross reactivity based on the sensors with immobilized LPS from Escherichia coli and Bacillus subtilis confirmed selectivity of this antibody. Furthermore, the 4H3B9D3 antibody was successfully isotypized as IgM using the piezoelectric sensors with secondary antibodies. Kinetics parameters of antibody were evaluated in the flow-through arrangement. The kinetic rate constants for the antibody 4H3B9D3 were ka = (2.31 ± 0.20)·105 l mol-1s-1 (association) and kd = (0.0010 ±0.00062) s-1 (dissociation) indicating very good affinity to the LPS antigen.

Pohanka, Miroslav; Pavlis, Oto; Skladal, Petr

2007-01-01

326

Transformation-Related Antigens Identified by Monoclonal Antibodies  

NASA Astrophysics Data System (ADS)

Tumor-cell proteins that were antigenic in a syngeneic animal were identified by immunoprecipitation with monoclonal antibodies. Spleen cells of BALB/c mice immunized with plasma membranes of Kirsten RNA sarcoma virus-transformed BALB/3T3 cells were fused with NS-l myeloma cells. Antibodies secreted into the culture fluid from these hybridomas were distinguished by their reactivity against proteins of different target cells. A total of 191 cultures were established; 143 produced antibodies that bound to BALB/3T3 cells transformed by the RNA sarcoma virus, of which antibodies from 82 bound to BALB/3T3 transformed with simian virus 40, and antibodies from 56 bound to BALB/3T3 cells. Thus, more than 50% of the cultures produced antibodies that possibly were specific to antigens of the transformed cell. Twenty different hybridomas have been cloned, and antibodies from eight of these were found to immunoprecipitate five different proteins. A protein of approximately 32,000 daltons was precipitated from BALB/3T3 cells transformed by the RNA sarcoma virus, simian virus 40, or methylcholanthrene but not from untransformed BALB/3T3 cells. A protein of about 300,000 daltons was precipitated from all four cell lines; precipitation was enhanced in the viral transformed cells. Proteins of approximately 57,000, 54,000, and 8500 daltons were immunoprecipitated from all four cell lines.

Strand, Mette

1980-06-01

327

A novel bispecific antibody recruits T cells to eradicate tumors in the "immunologically privileged" central nervous system  

PubMed Central

Bispecific T-cell engagers (BiTEs) may break multiple barriers that currently limit the use of immunotherapy in glioblastoma patients. We have recently described a novel BiTE specific for a mutated form of the epidermal growth factor receptor, EGFRvIII, that exerts potent antineoplastic effects against established invasive tumors of the brain.

Choi, Bryan D.; Pastan, Ira; Bigner, Darell D.; Sampson, John H.

2013-01-01

328

A novel bispecific antibody recruits T cells to eradicate tumors in the "immunologically privileged" central nervous system.  

PubMed

Bispecific T-cell engagers (BiTEs) may break multiple barriers that currently limit the use of immunotherapy in glioblastoma patients. We have recently described a novel BiTE specific for a mutated form of the epidermal growth factor receptor, EGFRvIII, that exerts potent antineoplastic effects against established invasive tumors of the brain. PMID:23734318

Choi, Bryan D; Pastan, Ira; Bigner, Darell D; Sampson, John H

2013-04-01

329

F(ab')2 fragment of a gp41 NHR-trimer-induced IgM monoclonal antibody neutralizes HIV-1 infection and blocks viral fusion by targeting the conserved gp41 pocket.  

PubMed

Using a recombinant protein N46FdFc that mimics the HIV-1 gp41 N-helix trimer to immunize mice, we identified the first IgM monoclonal antibody 18D3 that specifically bound to the conserved gp41 pocket. Its F(ab')2 fragment potently inhibited HIV-1 Env-mediated cell-cell fusion and neutralized infection by laboratory-adapted and primary HIV-1 isolates with different subtypes and tropism, including the T20-resistant variants. This F(ab')2 fragment can be used to develop a bispecific broad neutralizing monoclonal antibody or HIV-1 inactivator as a novel immunotherapeutic for treatment and prevention of HIV-1 infection. PMID:24141089

Lu, Lu; Wei, Meili; Chen, Yanxia; Xiong, Weiliang; Yu, Fei; Qi, Zhi; Jiang, Shibo; Pan, Chungen

2013-11-01

330

Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus  

Microsoft Academic Search

Neutralization of West Nile virus (WNV) in vivo correlates with the development of an antibody response against the viral envelope (E) protein. Using random mutagenesis and yeast surface display, we defined individual contact residues of 14 newly generated monoclonal antibodies against domain III of the WNV E protein. Monoclonal antibodies that strongly neutralized WNV localized to a surface patch on

Theodore Oliphant; Michael Engle; Grant E Nybakken; Chris Doane; Syd Johnson; Ling Huang; Sergey Gorlatov; Erin Mehlhop; Anantha Marri; Kyung Min Chung; Gregory D Ebel; Laura D Kramer; Daved H Fremont; Michael S Diamond

2005-01-01

331

Using monoclonal antibodies to prevent mucosal transmission of epidemic infectious diseases.  

PubMed Central

Passive immunization with antibodies has been shown to prevent a wide variety of diseases. Recent advances in monoclonal antibody technology are enabling the development of new methods for passive immunization of mucosal surfaces. Human monoclonal antibodies, produced rapidly, inexpensively, and in large quantities, may help prevent respiratory, diarrheal, and sexually transmitted diseases on a public health scale.

Zeitlin, L.; Cone, R. A.; Whaley, K. J.

1999-01-01

332

Production and characterisation of monoclonal antibodies to cell wall components of the flax rust fungus  

Microsoft Academic Search

Monoclonal antibodies have been raised against an haustorium-enriched sample prepared from flax leaves infected with the biotrophic flax rust pathogen Melampsora lini. The monoclonal antibodies were produced following conventional and co-immunisation procedures and the range of antibody specificities was compared. The preparation used as immunogen for the conventional protocol was a crude isolate of haustoria consisting of approx. 65% fungal

Leanne J. Murdoch; Issei Kobayashi; Adrienne R. Hardham

1998-01-01

333

Methods for the generation of chicken monoclonal antibody fragments by phage display  

Microsoft Academic Search

Phage display has become an important approach for the preparation of monoclonal antibodies from both immune and nonimmune sources. This approach allows for the rapid selection of monoclonal antibodies without the restraints of the conventional hybridoma approach. Although antibodies to a wide variety of antigens have been selected using phage display, some highly conserved mammalian antigens have proven to be

Jennifer Andris-Widhopf; Christoph Rader; Peter Steinberger; Roberta Fuller; Carlos F Barbas III

2000-01-01

334

Monoclonal antibody JC1: new reagent for studying cell proliferation.  

PubMed Central

AIM: To characterise a newly developed mouse monoclonal antibody JC1 which recognises a nuclear antigen present in proliferating cells in normal tissues and neoplastic lesions, and which is absent in resting cells. METHODS: The methodology was established using a representative range of frozen sections from normal tissues and from certain tumours which were immunostained with antibodies Ki67 and JC1. The molecular weight of the antigen recognised by JC1 was obtained by western blot analysis and this was compared with that of Ki67. IM-9 cell lysates containing Ki67 derived plasmids were also tested with JC1 antibody. RESULTS: Biochemical investigation indicated that the antigen recognised by JC1 gives two molecular weight bands of 212 and 123 kilodaltons, which is distinct from the well characterised anti-proliferation monoclonal antibody Ki67 (395-345 kilodaltons). In addition recombinant Ki67 protein is not recognised by JC1. Immunohistological reactivity was seen in areas known to contain numerous proliferating cells such as lymphoid germinal centres, splenic white matter, cortical thymocytes and undifferentiated spermatogonia. In tumours many cells from adenocarcinomas, oat cell carcinomas, squamous cell carcinomas of lung, and seminomas were labelled by JC1 with a distribution and proportion similar to that seen with Ki67. In normal tissues the only apparent difference was in testis where JC1 stained a considerably greater number of cells than Ki67. In all cases studied the new antibody showed nuclear reactivity only. JC1 did not show any cytoplasmic crossreactivity with squamous cells as is frequently seen with Ki67. CONCLUSION: Antibody JC1, which recognises a nuclear antigen present in proliferating cells, should provide a useful adjunct to Ki67 as a marker of proliferation especially in those cases such as squamous cell carcinomas where a Ki67 index cannot be determined. Images

Garrido, M C; Cordell, J L; Becker, M H; Key, G; Gerdes, J; Jones, M; Gatter, K C; Mason, D Y

1992-01-01

335

Use of heteropolymeric monoclonal antibodies to attach antigens to the C3b receptor of human erythrocytes: A potential therapeutic treatment  

SciTech Connect

The authors prepared bispecific, cross-linked monoclonal antibodies (heteropolymers) with specificity for both targeted antigens and the human erythrocyte (RBC) complement receptor. These heteropolymers facilitate binding of target antigens (human IgG and dinitrophenylated bovine {gamma} globulin) to human RBCs under conditions that either allow or preclude complement activation. Radioimmuno-assay analyses of this binding agree well with the number of complement receptors per RBC. In vitro whole-blood model experiments indicate heteropolymer-facilitated binding of antigens to RBCs is rapid and stable at 37C. It may be possible to extend these prototype experiments to the in vivo situation and use heteropolymer-attached RBCs for the safe and rapid binding, neutralization, and removal from the circulation of pathogenic antigens associated with infectious disease.

Taylor, R.P.; Sutherland, W.M.; Reist, C.J.; Webb, D.J.; Wright, E.L.; Labuguen, R.H. (Univ. of Virginia, Charlottesville (United States))

1991-04-15

336

Radiocurability by Targeting Tumor Necrosis Factor-{alpha} Using a Bispecific Antibody in Carcinoembryonic Antigen Transgenic Mice  

SciTech Connect

Purpose: Tumor necrosis factor-{alpha} (TNF-{alpha}) enhances radiotherapy (RT) killing of tumor cells in vitro and in vivo. To overcome systemic side effects, we used a bispecific antibody (BsAb) directed against carcinoembryonic antigen (CEA) and TNF-{alpha} to target this cytokine in a CEA-expressing colon carcinoma. We report the evaluation of this strategy in immunocompetent CEA-transgenic mice. Methods and Materials: The murine CEA-transfected colon carcinoma MC-38 was used for all experiments. In vitro, clonogenic assays were performed after RT alone, TNF-{alpha} alone, and RT plus TNF-{alpha}. In vivo, the mice were randomly assigned to treatment groups: control, TNF-{alpha}, BsAb, BsAb plus TNF-{alpha}, RT, RT plus TNF-{alpha}, and RT plus BsAb plus TNF-{alpha}. Measurements of endogenous TNF-{alpha} mRNA levels and evaluation of necrosis (histologic evaluation) were assessed per treatment group. Results: In vitro, combined RT plus TNF-{alpha} resulted in a significant decrease in the survival fraction at 2 Gy compared with RT alone (p < 0.00001). In vivo, we observed a complete response in 5 (50%) of 10, 2 (20%) of 10, 2 (18.2%) of 11, and 0 (0%) of 12 treated mice in the RT plus BsAb plus TNF-{alpha}, RT plus TNF-{alpha}, RT alone, and control groups, respectively. This difference was statistically significant when TNF-{alpha} was targeted with the BsAb (p = 0.03). The addition of exogenous TNF-{alpha} to RT significantly increased the endogenous TNF-{alpha} mRNA level, particularly when TNF-{alpha} was targeted with BsAb (p < 0.01). The percentages of necrotic area were significantly augmented in the RT plus BsAb plus TNF-{alpha} group. Conclusion: These results suggest that targeting TNF-{alpha} with the BsAb provokes RT curability in a CEA-expressing digestive tumor syngenic model and could be considered as a solid rationale for clinical trials.

Larbouret, Christel; Robert, Bruno [INSERM, Centre de Recherche en Cancerologie de Montpellier, Universite de Montpellier, and CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Linard, Christine [IRSN, Fontenay-aux-Roses (France); Teulon, Isabelle [INSERM, Centre de Recherche en Cancerologie de Montpellier, Universite de Montpellier, and CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Gourgou, Sophie M.Sc. [Unite de Biostatistiques en Oncologie, CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Bibeau, Frederic [Departement d'Anatomie Pathologique, CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Martineau, Pierre [Institut de Biotechnologie et Pharmacologie, CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Santoro, Lore [INSERM, Centre de Recherche en Cancerologie de Montpellier, Universite de Montpellier, and CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Departement d'Oncologie Radiotherapie, CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Pouget, Jean-Pierre; Pelegrin, Andre [INSERM, Centre de Recherche en Cancerologie de Montpellier, Universite de Montpellier, and CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Azria, David [INSERM, Centre de Recherche en Cancerologie de Montpellier, Universite de Montpellier, and CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France); Departement d'Oncologie Radiotherapie, CRLC Val d'Aurelle-Paul Lamarque, Montpellier (France)], E-mail: azria@valdorel.fnclcc.fr

2007-11-15

337

Epitope diversity of angiotensin II analysed with monoclonal antibodies.  

PubMed Central

The antigenic heterogeneity of angiotensin II (AII) was studied with monoclonal antibodies. Twelve antibodies were produced and characterized. Association constants for AII varied from 1.2 X 10(8) to 1.1 X 10(10) M-1. The fine specificity of the Mab was studied by immunoenzymoassay using solid-phase AII. Using AII analogues in binding inhibition experiments, three groups of specificity could be characterized: (1) five antibodies reacted only with peptides in which phenylalanine is the carboxy terminal aminoacid; for two of these antibodies, tyrosine4 is closely associated with the binding site, since iodine labelling suppresses reactivity; (2) two antibodies also required phenylalanine in position 8, but, in addition, reacted with AI, a decapeptide in which phenylalanine is not terminal; (3) five antibodies reacted with analogues in which phenylalanine had been substituted for another amino acid. In addition, studies in which binding of a biotinylated Mab to solid-phase AII was analysed in the presence of various unlabelled Mab suggest further antigenic heterogeneity of AII.

Picard, C; Ronco, P; Moullier, P; Yao, J; Baudouin, B; Geniteau Legendre, M; Verroust, P

1986-01-01

338

Characterization of a monoclonal anti-idiotype antibody to human anti-factor VIII antibodies.  

PubMed Central

Approximately 15% of individuals with hemophilia A develop antibodies (inhibitors) to therapeutically infused factor VIII that interfere with F.VIII coagulant activity. By using isoelectric focusing and immunospecific detection of anti-factor VIII antibodies, inhibitor plasma showed varied patterns of reactivity characteristic of a polyclonal response. Inhibitor plasma from patient Bt was observed to have an isolated banding pattern, or spectrotype, at pI 8.4 (SP8.4) distinct from his remaining anti-factor VIII antibodies. SP8.4 antibodies from this patient were partially purified and used to prepare monoclonal anti-idiotype antibodies. Monoclonal antibody Mab20-2H was found to detect a spectrotype in isoelectric-focused Bt plasma identical to SP8.4 and to bind anti-factor VIII antibodies. Furthermore, Mab20-2H binding could inhibit the binding of these anti-factor VIII antibodies to antigen, indicating that Mab20-2H recognizes an idiotope associated with antigen binding. Mab20-2H was also found to recognize antibodies from another inhibitor patient. This and other anti-idiotype reagents will be useful for defining genetic factors involved in the human immune response to factor VIII and in designing approaches to prevent or ameliorate this response. Images

Lubahn, B C; Reisner, H M

1990-01-01

339

Effects of monoclonal antibodies directed at cell surface molecules on murine experimental autoimmune uveoretinitis  

Microsoft Academic Search

· Background: To elucidate the immunopathogenic mechanism of endogenous uveitis, the effects of monoclonal antibodies to molecules\\u000a involved in the immune response were studied in murine experimental autoimmune uveoretinitis (EAU).?· Methods: Monoclonal\\u000a antibodies to CD4, CD8, Iak, Iad, lymphocyte function-associated antigen-1 (LFA-1), and intercellular adhesion molecule-1 (ICAM-1) were used in this study.\\u000a The monoclonal antibodies were added in the culture

Kazuhiko Ando; Yujiro Fujino; Manabu Mochizuki

1999-01-01

340

Identification of parietal cells in gastric body mucosa with HMFG-2 monoclonal antibody  

Microsoft Academic Search

AIMS--To identify parietal cells in the upper gastrointestinal tract by an immunoperoxidase method, using commercially available monoclonal antibodies. METHODS--Routine surgical biopsy specimens of gastric body mucosa were examined using the avidin-biotin peroxidase method with the monoclonal antibodies HMFG-1 and HMFG-2 to identify parietal cells. Double immunoperoxidase labelling with HK12.18, a well characterised monoclonal antibody directed against an epitope on the

M M Walker; A Smolka; J M Waller; D J Evans

1995-01-01

341

Monoclonal antibody therapy and renal transplantation: focus on adverse effects.  

PubMed

A series of monoclonal antibodies (mAbs) are commonly utilized in renal transplantation as induction therapy (a period of intense immunosuppression immediately before and following the implant of the allograft), to treat steroid-resistant acute rejections, to decrease the incidence and mitigate effects of delayed graft function, and to allow immunosuppressive minimization. Additionally, in the last few years, their use has been proposed for the treatment of chronic antibody-mediated rejection, a major cause of late renal allograft loss. Although the exact mechanism of immunosuppression and allograft tolerance with any of the currently used induction agents is not completely defined, the majority of these medications are targeted against specific CD proteins on the T or B cells surface (e.g., CD3, CD25, CD52). Moreover, some of them have different mechanisms of action. In particular, eculizumab, interrupting the complement pathway, is a new promising treatment tool for acute graft complications and for post-transplant hemolytic uremic syndrome. While it is clear their utility in renal transplantation, it is also unquestionable that by using these highly potent immunosuppressive agents, the body loses much of its innate ability to mount an adequate immune response, thereby increasing the risk of severe adverse effects (e.g., infections, malignancies, haematological complications). Therefore, it is extremely important for clinicians involved in renal transplantation to know the potential side effects of monoclonal antibodies in order to plan a correct therapeutic strategy minimizing/avoiding the onset and development of severe clinical complications. PMID:24590384

Zaza, Gianluigi; Tomei, Paola; Granata, Simona; Boschiero, Luigino; Lupo, Antonio

2014-03-01

342

Immunolocalization of neuroblastoma using radiolabeled monoclonal antibody UJ13A  

SciTech Connect

The monoclonal antibody UJ13A, raised after immunization of mice with human fetal brain, recognized an antigen expressed on human neuroblastoma cell lines and fresh tumors. Antibody was purified and radiolabeled with iodine isotopes using chloramine-T. In preclinical studies, 125I-labeled UJ13A was injected intravenously into nude mice bearing xenografts of human neuroblastoma. Radiolabeled UJ13A uptake by the tumors was four to 23 times greater than that by blood. In control animals, injected with a similar quantity of a monoclonal antibody known not to bind to neuroblastoma cells in vitro (FD44), there was no selective tumor uptake. Nine patients with histologically confirmed neuroblastoma each received 100 to 300 micrograms UJ13A radiolabeled with 1 to 2.8 mCi 123I or 131I. Sixteen positive sites were visible on gamma scans 1 to 7 days after injection: 15 were primary or secondary tumor sites, and one was a false positive; there were two false negatives. In two of the 15 positive sites, tumor had not been demonstrated by other imaging techniques; these were later confirmed as areas of malignant infiltration. No toxicity was encountered.

Goldman, A.; Vivian, G.; Gordon, I.; Pritchard, J.; Kemshead, J.

1984-08-01

343

Monoclonal antibodies for immunodectection of fibrin deposits on cancer cells.  

PubMed

The progression of a tumor from benign to malign and localized to invasive and metastatic growth is the major cause of poor outcome of therapy in cancer patients. The deposition of fibrin along with other pro-coagulant molecules into the extracellular matrix obviously serves as a scaffold to support proliferation, migration and tumor cell growth as well as protection against the immune system. The use of antibodies as agents for the immunodetection of fibrin deposits in vivo has been hampered by anti-fibrin cross-reactivities with fibrinogen. For the immunohistochemical detection of fibrin we used highly specific monoclonal antibodies to a synthetic fibrinunique peptide, because the fibrin molecule shares many epitopes with fibrinogen. The monoclonal antibody was applied to adenocarcinoma of colon, mamma, pancreas, sarcoma and acute myeloic leukemia. In all tissue sections and cytospin preparations fibrin was identified in a direct apposition to the surface membranes of carcinoma and sarcoma cells, predominantly at the host-tumor interface and also in regions directly adjacent to zones of angiogenesis, whereas normal cells and tissue showed no deposits of fibrin. The findings will be supported by investigations that factors and components of the coagulation system could be detected in the tumor stroma and tumor cells. These factors are obviously produced and secreted by the malignant cells and deposited together with fibrinogen into the extracellular matrix. Our results show that basically all malignant cells examined, independently of ectodermal or mesenchymal derivation, themselves are the origin of hypercoagulability and fibrinolytic system inhibition. PMID:23371915

Schardt, Friedrich W; Schmausser, Bernd; Bachmann, Eva

2013-08-01

344

Evaluation of oriented lysozyme immobilized with monoclonal antibody  

NASA Astrophysics Data System (ADS)

The orientation of a lysozyme immobilized with a monoclonal antibody was evaluated based on determination of the uppermost surface structure using time-of-flight secondary ion mass spectrometry (TOF-SIMS). Specific peaks of the oriented lysozyme immobilized with monoclonal anti-lysozyme antibody were obtained in comparison with reference samples, non-oriented immobilized lysozyme and immobilized anti-lysozyme antibody. All samples were freeze-dried before TOF-SIMS measurement, and then each sample was measured using TOF-SIMS with a bismuth cluster ion source. TOF-SIMS spectra were analyzed to select peaks specific to the oriented immobilized lysozyme as well as to identify their chemical formula and ensemble of amino acids. The possible chemical formulae of the lysozyme fragments were then investigated with an element matching program and a residue matching program. The results from TOF-SIMS spectra analysis were compared to the amino acid sequence of the lysozyme and its three-dimensional structure registered in the protein data bank. Finally, the fragment-ion-generating regions of the oriented immobilized lysozyme were determined based on the suggested residues and the three-dimensional structure.

Aoyagi, Satoka; Okada, Keigo; Shigyo, Ayako; Man, Naoki; Karen, Akiya

2008-12-01

345

Precipitation of a Monoclonal Antibody by Soluble Tungsten  

PubMed Central

Tungsten microparticles may be introduced into some pre-filled syringes during the creation of the needle hole. In turn, these microcontaminants may interact with protein therapeutics to produce visible particles. We found that soluble tungsten polyanions formed in acidic buffer below pH 6.0 can precipitate a monoclonal antibody within seconds. Soluble tungsten in pH 5.0 buffer at about 3 ppm was enough to cause precipitation of a mAb formulated at 0.02 mg/mL. The secondary structure of the protein was near-native in the collected precipitate. Our observations are consistent with the coagulation of a monoclonal antibody by tungsten polyanions. Tungsten-induced precipitation should only be a concern for proteins formulated below about pH 6.0 since tungsten polyanions are not formed at higher pHs. We speculate that the heterogenous nature of particle contamination within the poorly mixed syringe tip volume could mean that a specification for tungsten contamination based on the entire syringe volume is not appropriate. The potential potency of tungsten metal contamination is highlighted by the small number of particles that would be required to generate soluble tungsten levels needed to coagulate this antibody at pH 5.0.

Bee, Jared S.; Nelson, Stephanie A.; Freund, Erwin; Carpenter, John F.; Randolph, Theodore W.

2009-01-01

346

Practical considerations for nonclinical safety evaluation of therapeutic monoclonal antibodies  

PubMed Central

Monoclonal antibodies (mAbs) are a well established class of therapeutics as evidenced by a large number of FDA approved mAbs for the treatment of cancers and autoimmune diseases. Monoclonal antibodies that are molecularly engineered for enhanced functions and pharmacokinetic properties are routinely being considered for development by many biotechnology companies. Safety evaluation of current generation of mAbs poses new challenges due to the highly complex nature of engineering aspects and variability induced by the diverse recombinant cell systems to generate them. This review provides a basic outline for nonclinical safety evaluation of therapeutic antibodies. Important considerations for planning a preclinical program, the types of nonclinical safety studies, and a general timeline for their conduct in relation to clinical trials are described. A list of relevant regulatory documents issued by government agencies is also provided. Adoption of these principles will greatly enhance the quality and relevance of the nonclinical safety data generated and will facilitate future development of mAb therapeutics.

Lynch, Carmel M; Hart, Bruce W

2009-01-01

347

Characterization of oxidative carbonylation on recombinant monoclonal antibodies.  

PubMed

In the biotechnology industry, oxidative carbonylation as a post-translational modification of protein pharmaceuticals has not been studied in detail. Using Quality by Design (QbD) principles, understanding the impact of oxidative carbonylation on product quality of protein pharmaceuticals, particularly from a site-specific perspective, is critical. However, comprehensive identification of carbonylation sites has so far remained a very difficult analytical challenge for the industry. In this paper, we report for the first time the identification of specific carbonylation sites on recombinant monoclonal antibodies with a new analytical approach via derivatization with Girard's Reagent T (GRT) and subsequent peptide mapping with high-resolution mass spectrometry. Enhanced ionization efficiency and high quality MS(2) data resulted from GRT derivatization were observed as key benefits of this approach, which enabled direct identification of carbonylation sites without any fractionation or affinity enrichment steps. A simple data filtering process was also incorporated to significantly reduce false positive assignments. Sensitivity and efficiency of this approach were demonstrated by identification of carbonylation sites on both unstressed and oxidized antibody bulk drug substances. The applicability of this approach was further demonstrated by identification of 14 common carbonylation sites on three highly similar IgG1s. Our approach represents a significant improvement to the existing analytical methodologies and facilitates extended characterization of oxidative carbonylation on recombinant monoclonal antibodies and potentially other protein pharmaceuticals in the biotechnology industry. PMID:24731230

Yang, Yi; Stella, Cinzia; Wang, Weiru; Schöneich, Christian; Gennaro, Lynn

2014-05-20

348

A human fetal monoclonal DNA-binding antibody shares idiotypes with fetal and adult murine monoclonal DNA-binding antibodies.  

PubMed Central

A human DNA-binding monoclonal antibody was produced by fusing the hepatocytes from a 12-week-old human fetus with the lymphoblastoid cell line GM 4672 using polyethylene glycol. This antibody, designated BEG 2, binds to single-stranded (ss) DNA but also binds to double-stranded (ds) DNA, poly(dT), polyI and poly(ADP-ribose), but not to RNA, cardiolipin or K-30. The binding of BEG 2 to these polynucleotides can be inhibited by incubation with polynucleotides in the fluid phase. A rabbit polyclonal anti-idiotype was raised, and using this reagent it was shown that the BEG 2 idiotype is present in normal human serum (7%), systemic lupus erythematosus (SLE) sera (8%) and rheumatoid arthritis sera (23%). The extent of idiotypic sharing between BEG 2 and murine monoclonal DNA-binding antibodies, in particular monoclonal antibody (mAb) 423 (derived from a 15-day-old fetal MRL/Mp-lpr/lpr mouse) and mAb 402 (derived from an adult MRL/lpr mouse), was also investigated. Using a competition ELISA, it was shown that preincubation of BEG 2 with rabbit anti-423 and rabbit anti-402 inhibits the binding of BEG 2 to DNA, and the binding of 402 to DNA by anti-BEG 2 and anti-423. These data suggest that mAb BEG 2, 423 and 402 share common idiotypes, that autoreactivity is present in early fetal life, and that autoantibodies may be encoded for by germline genes, which have been conserved through evolution. Images Figure 2 Figure 3 Figure 7

Watts, R A; Ravirajan, C T; Staines, N A; Isenberg, D A

1990-01-01

349

Detecting low level sequence variants in recombinant monoclonal antibodies.  

PubMed

A systematic analytical approach combining tryptic and chymotryptic peptide mapping with a Mascot Error Tolerant Search (ETS) has been developed to detect and identify low level protein sequence variants, i.e., amino acid substitutions, in recombinant monoclonal antibodies. The reversed-phase HPLC separation with ultraviolet (UV) detection and mass spectral acquisition parameters of the peptide mapping methods were optimized by using a series of model samples that contained low levels (0.5-5.0%) of recombinant humanized anti-HER2 antibody (rhumAb HER2) along with another unrelated recombinant humanized monoclonal antibody (rhumAb A). This systematic approach's application in protein sequence variant analysis depends upon time and sensitivity constraints. An example of using this approach as a rapid screening assay is described in the first case study. For stable CHO clone selection for an early stage antibody project, comparison of peptide map UV profiles from the top four clone-derived rhumAb B samples quickly detected two sequence variants (M83R at 5% and P274T at 42% protein levels) from two clones among the four. The second case study described in this work demonstrates how this approach can be applied to late stage antibody projects. A sequence variant, L413Q, present at 0.3% relative to the expected sequence of rhumAb C was identified by a Mascot-ETS for one out of four top producers. The incorporation of this systematic sequence variant analysis into clone selection and the peptide mapping procedure described herein have practical applications for the biotechnology industry, including possible detection of polymorphisms in endogenous proteins. PMID:20400866

Yang, Yi; Strahan, Alex; Li, Charlene; Shen, Amy; Liu, Hongbin; Ouyang, Jun; Katta, Viswanatham; Francissen, Kathleen; Zhang, Boyan

2010-01-01

350

Production of monoclonal antibody to acaricide dicofol and its derivatives.  

PubMed

In Thailand detection of acaricide dicofol residues has been sporadically performed due to the limitation of analytical techniques. Conventional analytical methods for detecting dicofol residues most often use chromatographic-based techniques. Our ultimate aim is to develop an alternative method for rapidly analyzing dicofol residues in vegetables and fruit samples. Here we report the production of monoclonal antibodies specific to dicofol and its derivatives. Hapten-protein carriers were prepared by linking succinic anhydride to dichlorobenzhydrol (DCBH), which was then conjugated to bovine serum albumin (BSA) and oval albumin (OVA). DCBH-BSA conjugate was used as immunogen while DCBH-OVA conjugate was used as capture antigen for competitive inhibition assay. Female BALB/c mice were immunized with DCBH-BSA conjugate subcutaneously, and antibody (Ab) level was determined 2 weeks after the last immunization. Spleen cells producing high titer antibody were isolated and fused with myeloma cells of P3.X6.Ag8.653. After limiting dilutions, antibody produced by one clone had high affinity, which was found to be of IgG1 with ? light chain. Specificity and inhibition concentrations of the monoclonal antibody (MAb) were determined by competitive indirect ELISA with dicofol, and its 50% (IC(50)) was 0.28 ?g/mL. Working ranges of the developed immunoassay were from 0.07 to 25 ?g/mL. Hence, the prepared MAb will be able to be applied for immunoassay development for detecting dicofol residue in vegetables and fruits far below the maximum residue limit such that 5 g of fruits and berries can be detected below 0.1 mg/kg. PMID:21118018

Hongsibsong, Surat; Prapamontol, Tippawan; Suphavilai, Chaisuree; Wipasa, Jiraprapa; Pattarawarapan, Mookda; Kasinrerk, Watchara

2010-12-01

351

Production and characterization of monoclonal antibodies specific for Mycobacterium bovis.  

PubMed

A series of monoclonal antibodies (MAbs), specific for Mycobacterium bovis and BCG strains, were tested extensively for cross-reactivity to a wide range of mycobacterial species using ELISA, Western blotting and dot-blot analysis. The MAbs bound specifically to M. bovis and BCG and showed limited cross-reactivity with some strains of M. tuberculosis. All these MAbs recognized a 22 kDa protein previously termed MPB70, and by competitive ELISA analysis appeared to detect at least three M. bovis-specific determinants on the MPB70 molecule. PMID:2474629

Wood, P R; Ripper, J; Radford, A J; Bundesen, P G; Rylatt, D B; Cottis, L E; John, M; Plackett, P

1988-09-01

352

Positron emission tomographic imaging of tumors using monoclonal antibodies  

SciTech Connect

The overall objective of this research project is to develop methods for utilizing positron emission tomography (PET) to increase the clinical potential of radiolabeled monoclonal antibodies (MAbs). Both diagnostic and therapeutic applications of labeled MAbs could be improved as a result of knowledge obtained through the exploitation of the advantageous imaging characteristics associated with PET. By labeling MAbs with positron-emitting nuclides, it should be possible to quantitate the dynamics of their three-dimensional distribution in vivo. Our long-term goals are to apply this approach. 3 tabs.

Zalutsky, M.R. (Duke Univ., Durham, NC (USA). Dept. of Radiology)

1989-12-01

353

Drug evaluation: tefibazumab--a monoclonal antibody against staphylococcal infection.  

PubMed

Inhibitex Inc is investigating tefibazumab, a humanized monoclonal antibody specific for the fibrin-binding surface epitope clumping factor A protein (expressed on the surface of most Staphylococcus aureus strains), for the potential intravenous prevention and/or treatment of S aureus infections. In June 2006, Inhibitex was seeking to outlicense certain development rights to the drug. Pending the outcome of partnering discussions, Inhibitex suspended the initiation of any additional clinical trials of tefibazumab. Preclinical in vivo studies were ongoing at that time. PMID:17078388

John, Joseph F

2006-10-01

354

Generation of cell lines for monoclonal antibody production.  

PubMed

Monoclonal antibodies (mAbs) represent the largest group of therapeutic proteins with 30 products approved in the USA and hundreds of therapies currently undergoing clinical trials. The complex nature of mAbs makes their development as therapeutic agents constrained by numerous criteria such as quality, safety, regulation, and quantity. Identification of a clonal cell line expressing high levels of mAb with adequate quality attributes and generated in compliance with regulatory standards is a necessary step prior to a program moving to large-scale production for clinical material. This chapter outlines the stable transfection technology that generates clonal cell lines for commercial manufacturing processes. PMID:24515472

Alvin, Krista; Ye, Jianxin

2014-01-01

355

Production and characterization of mouse monoclonal antibodies against Afipia felis.  

PubMed

A series of 10 monoclonal antibodies reacting with Afipia felis antigens were selected from mice immunized with live organisms of the reference strain ATCC 53690. Immunoblotting against SDS-PAGE-separated A felis sonicate allowed the antibodies to be classified into three groups: 1) 168-4, -6, -7 and -10 reacted with a 53 kDa antigen, 2) 168-1, -3 and -9 reacted with both 53 kDa and 60 kDa antigens, and 3) 168-2, -5 and -9 reacted with other antigens. Antibodies of group 1 did not cross-react with other Afipia species or 36 unrelated bacteria, whereas those of groups 2 and 3 reacted with other Afipia species and some unrelated bacteria. Immunoblots of crossed immunoelectrophoretic patterns of A. felis sonicate against rabbit antiserum showed that antibodies of groups 1 and 2 bound to the same precipitin arcs. Antibodies of group 1 reacted with a species-specific epitope on the 53 kDa antigen, while those of group 2 reacted with other epitopes shared by the 53 kDa and 60 kDa antigens. The binding of antibodies of group 1 to A. felis sonicate was inhibited by post-infection rabbit serum, whereas no inhibition was observed for antibodies of group 2. The species-specific epitope of the 53 kDa antigen and the early appearance of antibodies against this epitope after infection suggest that this antigen can be used in a serodiagnostic test for A. felis infection. PMID:9137514

Engbaek, K; Uttenthal, L O; Koch, C

1997-03-01

356

Preparation and characterization of a monoclonal antibody with specific affinity for pyrimidine dimers in RNA  

Microsoft Academic Search

A monoclonal antibody was prepared which shows specific affinity for pyrimidine dimers in RNA. The antibody meets three criteria for pyrimidine dimer-dependent affinity. (1) Antibody binding is dependent upon UV irradiation of nucleic acids. Irradiation both at 313 nm in the presence of a sensitizer and at 270 nm without sensitizer promotes antibody binding to RNA and polyribonucleotides. (2) Antibody

1986-01-01

357

A novel bispecific tetravalent antibody fusion protein to target costimulatory activity for T-cell activation to tumor cells overexpressing ErbB2/HER2.  

PubMed

Persistent activation of T-lymphocytes requires two signals: one is initiated by T-cell receptor binding to antigenic peptide presented by MHC molecules. In addition, binding of the B7 family members CD80 or CD86 on professional antigen presenting cells to CD28 on T cells is considered to provide an important costimulatory signal. Activation without costimulation induces T-cell unresponsiveness or anergy. To selectively localize costimulatory activity to the surface of tumor cells and enhance activation of tumor-specific T cells, we have developed a novel molecular design for bispecific costimulatory proteins with antibody-like structure. Within a single polypeptide chain we have assembled the IgV-like, CD28-binding domain of human CD86 (CD86(111)) together with hinge, CH2 and CH3 domains of human IgG1, and the scFv(FRP5) antibody fragment which recognizes the ErbB2 (HER2) protooncogene present at high levels on the surface of many human tumor cells. Upon expression in the yeast Pichia pastoris, the resulting CD86(111)-IgG-scFv(FRP5) protein could be purified as a homodimeric, tetravalent molecule from culture supernatants using single-step affinity chromatography. Bispecific binding of the molecule to ErbB2 on the surface of tumor cells and to the B7 counter receptor CTLA-4 was demonstrated by FACS analysis. Potent costimulatory activity of chimeric CD86(111)-IgG-scFv(FRP5) was confirmed by its ability to stimulate the proliferation of primary human lymphocytes pre-activated by low concentrations of anti-CD3 antibody. Our results suggest that such multivalent soluble proteins which combine specific targeting to tumor cells with costimulatory activity may become useful tools to elicit and/or improve T-cell mediated, tumor-specific immune responses. PMID:15713482

Biburger, Markus; Weth, Robert; Wels, Winfried S

2005-03-11

358

Emerging Therapies: Spectrum of Applications of Monoclonal Antibody Therapy.  

PubMed

This article focuses on the recent dramatic advances in the applications of monoclonal antibody therapy to hematopoietic and neoplastic disease. The increase in the understanding of the role of growth factors and their receptors in the pathogenesis of malignancy and other undesirable hematological events taken in conjunction with the ability to produce humanized chimeric monoclonal antibodies to these targets is providing a new perspective for the treatment of leukemia, lymphoma and breast cancer, autoimmune disease and for prevention of ischemic complications. Dr. Waldmann describes approaches targeting the Her2/neu and the II-2/IL-15 receptor systems. The Her2/neu receptor is overexpressed in select breast, ovarian, gastric and pancreatic neoplasms. The use of trastuzumab (Herceptin) in the treatment of patients with breast cancer whose tumors overexpress this receptor are reviewed. The IL-2 receptor (Tac) is expressed on select malignant cells (adult T cell leukemia, hairy cell leukemia) and activated T cells involved in autoimmune disease and organ rejection. Humanized anti-Tac alone (daclizumab, Zenapax) or armed with toxins or radionuclides have been used successfully in the treatment of leukemia. Dr. Levy updates the experience with rituximab targeting CD20 on B cell lymphomas and reviews the antibodies to CD3, CD22, CD33, CD52, HLA-DR beta chain and HLA-D currently in or proposed for clinical trials, including radiolabelled antibodies. In the last section, Dr. Coller reviews the therapeutic results achieved with abciximab (ReoPro), an antagonist of platelet receptor GPIIbIIIa for the prevention of restenosis in percutaneous coronary interventions and the treatment of unstable angina. The mechanism of action, pharmacology and safety and efficacy of abciximab are reviewed. PMID:11701553

Waldmann, Thomas A.; Levy, Ronald; Coller, Barry S.

2000-01-01

359

SPECT assay of radiolabeled monoclonal antibodies. Third yearly progress report, September 1991--February 1992.  

National Technical Information Service (NTIS)

The accurate determination of the biodistribution of radiolabeled monoclonal antibodies (MoAbs) is important for calculation of dosimetry and evaluation of pharmacokinetic variables such as antibody dose and route of administration. The hypothesis of this...

R. J. Jaszczak

1992-01-01

360

Characterization of monoclonal antibody size variants containing extra light chains  

PubMed Central

Size exclusion chromatography (SEC) is the most commonly used method to separate and quantify monoclonal antibody (mAb) size variants. MAb-A is an IgG1 subtype humanized monoclonal antibody recombinantly produced in Chinese hamster ovary (CHO) cells. SEC analysis of MAb-A resolved a peak, named Peak 1, which elutes between monomer and dimer peaks. MAb-A lots produced from different clones and production scales all have 0.2–0.3% of SEC Peak 1. Electron spray ionization—time of flight mass spectrometry (ESI-TOF MS), microfluidics capillary electrophoresis and sodium dodecyl sulfate-PAGE (SDS PAGE) results demonstrated that SEC Peak 1 contains two structural variants: MAb-A with one extra light chain (2H3L) and MAb-A with two extra light chains (2H4L). The C-terminal Cys of the extra light chain in Peak 1 variants is either a free thiol, capped by glutathione, cysteine, or another light chain. Both electrophoresis and LC/MS analyses of non-reduced and reduced samples suggested that the extra light chains are linked to the MAb-A light chain through disulfide bonds. Isolated SEC Peak 1 fraction had a potency of 50% relative to MAb-A reference material. The 50% potency loss may result from the reduced accessibility to the antigen-binding site caused by the extra light chain(s)’ steric hindrance.

Lu, Connie; Liu, Dandan; Liu, Hongbin; Motchnik, Paul

2013-01-01

361

Detection of grass carp reovirus (GCRV) with monoclonal antibodies.  

PubMed

Grass carp reovirus (GCRV) is a pathogen that causes hemorrhagic disease of grass carp. It is the most serious infectious disease of carp and causes serious losses of fingerlings of grass carp and black carp. In this study, a recombinant VP4, one of the viral core proteins, was constructed with a histidine tag and expressed at a high level in E. coli, and the expressed protein was mainly found in the form of inclusion bodies. The expressed VP4 protein was recognized by an anti-His-tag monoclonal antibody and goat anti-GCRV serum. Four monoclonal antibodies (16B7, 39E12, 13C3 and 14D1) against the recombinant VP4 protein were produced. These MAbs did not react with any of the tested viruses or fish cells lines in the ELISA tests except GCRV. In western blotting analysis, a protein band was observed when the recombinant VP4 protein of GCRV was used as an antigen, but a 68-kDa band was observed when natural capsid proteins of GCRV were used as antigens. Furthermore, a sandwich ELISA was developed for detection of GCRV. The detection limit of the test was 105 TCID50 of GCRV per mL. PMID:24122108

Hongli, Jing; Lifeng, Zhang; Zhenzhen, Fang; Lipu, Xu; Min, Zhang; Na, Wang; Yulin, Jiang; Xiangmei, Lin

2014-04-01

362

Monoclonal antibodies: a target therapy for multiple sclerosis.  

PubMed

Multiple sclerosis (MS) is a complex autoimmune disease of the central nervous system. It is characterized by a proinflammatory and neurodegenerative process that results in neuroaxonal damage. Over the last two decades, a wide range of immunomodulatory and immunosuppressive treatments have been used for the management of MS. Several treatments have been developed or are under evaluation for reducing relapses, disease progression and long-term MSrelated disability. Recently, a growing interest has emerged for therapeutics with very selective actions, particularly monoclonal antibodies, to target several biological pathways involved in MS. To date, only Natalizumab (Tysabri(®)) has been approved for the treatment of active MS forms. Its therapeutic mechanism is the blockade of the a4-integrin molecule of many leukocytes, which leads to a decrease of immune cells migration, in particular of lymphocytes, across the blood-brain barrier. Furthermore, other promising molecules are under study in clinical trials. In this review, we summarize and discuss the history, pharmacodynamics and safety of monoclonal antibodies that have been approved or are under evaluation for the selective treatment of MS. PMID:24479836

Lorefice, Lorena; Fenu, Giuseppe; Frau, Jessica; Coghe, Giancarlo; Marrosu, Maria Giovanna

2014-01-01

363

Bothropic antivenom based on monoclonal antibodies, is it possible?  

PubMed

Neutralizing monoclonal antibodies against three major toxic components of Bothrops atrox venom were produced and tested. The mAbs against phospholipase A2, hemorrhagic metalloprotease, and thrombin-like enzymes were produced in large amounts and purified with caprylic acid followed by ammonium sulfate precipitation. Purified mAbs were analyzed by SDS-PAGE and their ability to neutralize the respective toxins was tested. Five Swiss mice were injected i.p. with 13.5 mg of pooled mAbs and challenged via s.c. route with venom. Survival rate was recorded for the next 48 h. All mice treated and challenged with venom survived, whereas only one mouse in the control group survived. Bleeding time in mice treated with mAbs was similar to that observed in control mice. Our results show that monoclonal antibodies neutralized the lethal toxicity of Bothrops venom and indicate that there is a reasonable possibility of developing antivenoms based on humanized mAbs to treat victims of venomous animals in the future. PMID:23732123

Frauches, Thiago S; Petretski, Jorge H; Arnholdt, Andrea C V; Lasunskaia, Elena B; de Carvalho, Eulógio C Q; Kipnis, Thereza L; da Silva, Wilmar D; Kanashiro, Milton M

2013-09-01

364

Evidence for polyreactivity seen with monoclonal antibodies produced against type II collagen.  

PubMed

Specific type II collagen monoclonal antibodies are needed for the quantification of articular cartilage collagen. In this study we produced and characterized 29 type II collagen monoclonal antibodies. Hybridomas were generated from mice immunized with rat type II collagen, selected for high antibody production against type II collagen using ELISA. Antibodies from selected and cloned hybridoma cells were purified by affinity chromatography and their reactivity tested by ELISA against a panel of antigens including actin, thyroglobulin, and single stranded DNA, all of which have been used to characterize the 'naturally occurring antibodies.' It was found that many of the anti-type II collagen monoclonal antibodies reacted to more than one antigen. The monospecific antibodies had higher affinity to type II collagen than the antibodies which demonstrated multireactivity. Because of the prevalence of polyreactive anti-type II collagen antibodies, it is advisable to employ highly selective methodologies to isolate high affinity monospecific antibodies to type II collagen. PMID:2066572

Chichester, C; Barrach, H J; Chichester, A; Matoney, A; Srinivas, G R

1991-07-01

365

Intracavitary use of two radiolabeled tumor-associated monoclonal antibodies  

SciTech Connect

Six patients with metastatic breast cancer and malignant pleural effusions and 13 patients with known or suspected ovarian cancer, underwent immunoscintigraphy after intracavitary (intrapleural or intraperitoneal) administration of iodine-131-(131I) or indium-111-(111In) labeled tumor associated monoclonal antibodies HMFG2 and H17E2. This method proved to be sensitive and specific with a true-positive result in 13 out of 14 patients with tumor and a true-negative result in five out of five patients without tumor. At any one time, 65%-80% of the whole-body radioactivity was closely associated with the cavity into which the radiolabeled antibody was administered while the radioactivity in the blood was always low, (approximately 4 X 10(-3) of administered dose/ml of blood). Concentrations of radiolabeled antibody (per gram of tumor tissue) ranged from 0.02%-0.1% of the injected dose in intracavitary tumors, but only 0.002% in a retroperitoneal metastasis. The specificity of this approach was documented in four control patients with benign ovarian cysts and in two patients who were imaged using both specific and nonspecific radiolabeled antibody. We conclude that the intracavitary administration of 131I- or 111In-labeled HMFG2 and H17E2 is a favorable route of administration and offers significant advantages over previously reported intravenous administration for the localization of breast or ovarian metastases confined to the pleural or peritoneal cavities.

Malamitsi, J.; Skarlos, D.; Fotiou, S.; Papakostas, P.; Aravantinos, G.; Vassilarou, D.; Taylor-Papadimitriou, J.; Koutoulidis, K.; Hooker, G.; Snook, D.

1988-12-01

366

[Production and characteristics of monoclonal antibodies to the diphtheria toxin].  

PubMed

Monoclonal antibodies to the diphtheria toxin were produced without cross reactivity with the thermolabile toxin (LT) from Escherichia coli; ricin; choleraic toxin; the SeA, SeB, SeE, SeI, and SeG toxins of staphylococcus; the lethal factor of the anthrax toxin; and the protective antigen of the anthrax toxin. A pair of antibodies for the quantitative determination of the diphtheria toxin in the sandwich variation of enzyme-linked immunosorbent assay (ELISA) was chosen. The determination limit of the toxin was 0.7 ng/ml in plate and 1.6 ng/ml in microchip ELISA. The presence of a secretion from the nasopharynx lavage did not decrease the sensitivity of the toxin determination by sandwich ELISA. The immunization of mice with the diphtheria toxin and with a conjugate of the diphtheria toxin with polystyrene microspheres demonstrated that the conjugate immunization resulted in the formation of hybridoma clones which produced antibodies only to the epitopes of the A fragment of the diphtheria toxin. The immunization with the native toxin caused the production of hybridoma clones which predominantly produced antibodies to the epitopes of the B fragment. PMID:19915639

Valiakina, T I; Lakhtina, O E; Komaleva, R L; Simonova, M A; Samokhvalova, L V; Shoshina, N S; Kalinina, N A; Rubina, A Iu; Filippova, M A; Vertiev, Iu V; Grishin, E V

2009-01-01

367

Characterization of a monoclonal antibody to thymidine glycol monophosphate  

SciTech Connect

A monoclonal antibody specific for thymine glycol (TG) in irradiated or OsO4-treated DNA was obtained by immunizing with thymidine glycol monophosphate (TMP-glycol) conjugated to bovine serum albumin by a carbodiimide procedure. Screening by dot-immunobinding and enzyme-linked immunosorbant assay (ELISA) procedures gave eight clones that bound OsO4- treated DNA. One of them, 2.6F.6B.6C, an IgG2a kappa, was characterized further. Hapten inhibition studies with OsO4-treated DNA showed that the antibody was specific for TMP-glycol. Among the various inhibitors tested, inhibition was in the order TMP-glycol greater than 5,6-dihydrothymidine phosphate greater than TMP greater than thymidine glycol greater than TG. Inhibition by 5,6-dihydrothymidine, thymidine, thymine, AMP, and CMP was negligible. In OsO4-treated DNA, as few as 0.5 TG per 10,000 bp were detectable by direct ELISA. Inhibition assays could detect as few as 1.5 TG per 10,000 bp. The antibody was equally reactive with native or denatured DNA containing TG. Among the X-irradiated homopolymers dC, dA, dG, and dT, only dT reacted with the antibody. Using an ELISA, the antibody could detect damage in irradiated DNA at the level of 20 Gy. Thus the antibody is of potential use in assays for DNA damage caused by X rays or other agents that damage DNA by free radical interactions.

Chen, B.X.; Hubbard, K.; Ide, H.; Wallace, S.S.; Erlanger, B.F. (Columbia Univ. College of Physicians and Surgeons, New York, NY (USA))

1990-11-01

368

Polyclonal and monoclonal antibody therapy for experimental Pseudomonas aeruginosa pneumonia.  

PubMed Central

A human immunoglobulin G preparation, enriched in antibodies to lipopolysaccharide (LPS) Pseudomonas aeruginosa antigens (PA-IGIV) and murine monoclonal antibodies (MAb) to P. aeruginosa Fisher immunotype-1 (IT-1) LPS antigen and outer membrane protein F (porin), were evaluated for therapeutic efficacy in a guinea pig model of P. aeruginosa pneumonia. The concentration of antibodies to IT-1 LPS was 7.6 micrograms/ml in PA-IGIV and 478 micrograms/ml in the IT-1 MAb preparation. No antibody to IT-1 was detected in MAb to porin. For study, animals were infected by intratracheal instillation of IT-1 P. aeruginosa and then treated 2 h later with intravenous infusions of PA-IGIV, IT-1 MAb, or porin MAb. Control groups received intravenous albumin, and routinely died from pneumonia. Both PA-IGIV (500 mg/kg) and IT-1 MAb (greater than or equal to 2.5 mg/kg) treatment resulted in increased survival (P less than 0.01 to 0.001), and also improved intrapulmonary killing of bacteria. Porin MAb failed to protect from fatal pneumonia. IT-1 MAb treatment produced more survivals than did PA-IGIV treatment but only at dosages of MAb resulting in serum antibody concentrations greater than those achieved with PA-IGIV. PA-IGIV and IT-1 MAb demonstrated in vitro and in vivo (posttreatment guinea pig serum) opsonophagocytic activity for the IT-1 challenge strain. However, the polyclonal preparation required complement, whereas the MAb did not. We conclude that passive immunization with polyclonal hyperimmune P. aeruginosa globulin or with MAb to LPS antigens may be useful in the treatment of acute P. aeruginosa pneumonia. The relative efficacies of such preparations may be limited, however, by their type-specific LPS antibody concentrations.

Pennington, J E; Small, G J; Lostrom, M E; Pier, G B

1986-01-01

369

Scintigraphy of normal mouse ovaries with monoclonal antibodies to ZP-2, the major zona pellucida protein  

SciTech Connect

The zona pellucida is an extracellular glycocalyx, made of three sulfated glycoproteins, that surrounds mammalian oocytes. Parenterally administered monoclonal antibodies specific for ZP-2, the most abundant zona protein, localize in the zona pellucida. When labeled with iodine-125, these monoclonal antibodies demonstrate a remarkably high target-to-nontarget tissue ratio and provide clear external radioimaging of ovarian tissue.

East, I.J.; Keenan, A.M.; Larson, S.M.; Dean, J.

1984-08-31

370

Monoclonal antibodies in the detection of bone marrow metastases in small cell lung cancer  

Microsoft Academic Search

Using conventional examination (CE) of H&E stained slides from bone marrow aspirates, metastases can be detected in approximately 25% of patients with small cell lung cancer. We investigated a panel of monoclonal antibodies using immunohistochemistry in the diagnosis of bone marrow infiltration from SCLC and compared the results with CE. Seven monoclonal antibodies raised against epithelial antigens (CAM 5.2, MOV

BG Skov; FR Hirsch; L Bobrow

1992-01-01

371

Characterization of a monoclonal antibody with specificity for holo-transcobalamin  

Microsoft Academic Search

BACKGROUND: Holotranscobalamin, cobalamin-saturated transcobalamin, is the minor fraction of circulating cobalamin (vitamin B12), which is available for cellular uptake and hence is physiologically relevant. Currently, no method allows simple, direct quantification of holotranscobalamin. We now report on the identification and characterization of a monoclonal antibody with a unique specificity for holotranscobalamin. METHODS: The specificity and affinity of the monoclonal antibodies

Lars Orning; Anne Rian; Andrew Campbell; Jeff Brady; Sergey N Fedosov; Birgit Bramlage; Keith Thompson; Edward V Quadros

2006-01-01

372

Differentiation of F38 mycoplasmas causing contagious caprine pleuropneumonia with a growth-inhibiting monoclonal antibody.  

PubMed Central

Monoclonal antibody WM-25 inhibited the in vitro growth of 13 F38 isolates from goats with contagious caprine pleuropneumonia but not 7 heterologous mycoplasma isolates representing four different species. In contrast to results with polyclonal antisera, growth inhibition by monoclonal antibody WM-25 was specific for F38 mycoplasma isolates and constituted a reliable means of distinguishing F38 from other mycoplasmas.

Rurangirwa, F R; McGuire, T C; Musoke, A J; Kibor, A

1987-01-01

373

Monoclonal antibodies to cyclodiene insecticides and method for detecting the same  

DOEpatents

Methods are described for making specific monoclonal antibodies useful for detection of cyclodienes in foods and environmental samples. Monoclonal antibodies specifically reactive with cyclodienes can detect accumulated pesticides in food, tissue or environmental samples. Extraction and preparation of organic samples for immunoassay in a polar-nonpolar reaction medium permits detection of halogenated organic ring structures at concentrations in samples.

Stanker, Larry H. (Livermore, CA); Vanderlaan, Martin (Danville, CA); Watkins, Bruce E. (Livermore, CA)

1994-01-01

374

Monoclonal antibodies to cyclodiene insecticides and method for detecting the same  

DOEpatents

Methods are described for making specific monoclonal antibodies useful for detection of cyclodienes in foods and environmental samples. Monoclonal antibodies specifically reactive with cyclodienes can detect accumulated pesticides in food, tissue or environmental samples. Extraction and preparation of organic samples for immunoassay in a polar-nonpolar reaction medium permits detection of halogenated organic ring structures at concentrations in samples. 13 figs.

Stanker, L.H.; Vanderlaan, M.; Watkins, B.E.

1994-08-02

375

Isolation and characterization of a monoclonal antibody directed against type 1 fimbriae organelles from Escherichia coli.  

PubMed Central

We have isolated a mouse immunoglobulin M (IgM) monoclonal antibody directed against type 1 fimbriae from the Escherichia coli K-12-derived strain CSH50. Antibody specificity was demonstrated by (i) the ability of fimbriate but not nonfimbriate bacteria to compete with solid-phase purified fimbriae for antibody binding in an enzyme-linked immunosorbent assay, (ii) the visualized binding of antibody to fimbriae alone by electron microscopy, and (iii) the appearance in a radioimmunoprecipitation assay of a single electrophoretic band comigrating with pure type 1 fimbriae. The monoclonal antibody was further characterized by immunoblot analysis and compared with previously prepared rabbit anti-fimbrial antisera. Whereas the monospecific but polyvalent antisera recognized both fimbrial monomeric subunits and non-disaggregated fimbriae organelles, the monoclonal antibody recognized only the intact organelles even when the samples were prepared under nondenaturing conditions. The monoclonal antibody, therefore, might be directed against an epitope spanning two (or more) adjacent fimbrial subunits. Images

Eisenstein, B I; Clements, J R; Dodd, D C

1983-01-01

376

Identification of hog cholera viral isolates by use of monoclonal antibodies to pestiviruses.  

PubMed

A collection of 90 field isolates of hog cholera virus (HCV) was used to test the specificity of four hybridoma cell lines secreting monoclonal antibodies against pestiviruses. Reaction of virus isolates and monoclonal antibodies was controlled by an indirect immunofluorescence assay (IFA). Two monoclonal antibodies which had been generated against HC virus strain "Alfort 187" were reactive only with HCV field isolates and an HCV reference strain but not with bovine viral diarrhoea virus (BVDV) reference strains. Two other monoclonal antibodies (generated against BVDV, strain NADL) reacted only with BVDV reference strains but not with HCV field isolates, although with 3 of these strains focal reactions involving only a few cells were detected. The ability to discriminate between both viruses is a diagnostic need which may be fulfilled by these monoclonal antibodies. PMID:2837867

Hess, R G; Coulibaly, C O; Greiser-Wilke, I; Moennig, V; Liess, B

1988-04-01

377

Selection of Ceratitis capitata (Diptera: Tephritidae) Specific Recombinant Monoclonal Phage Display Antibodies for Prey Detection Analysis  

PubMed Central

Several recombinant antibodies against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), one of the most important pests in agriculture worldwide, were selected for the first time from a commercial phage display library of human scFv antibodies. The specificity and sensitivity of the selected recombinant antibodies were compared with that of a rabbit polyclonal serum raised in parallel using a wide range of arthropod species as controls. The selected recombinant monoclonal antibodies had a similar or greater specificity when compared with classical monoclonal antibodies. The selected recombinant antibodies were successfully used to detect the target antigen in the gut of predators and the scFv antibodies were sequenced and compared. These results demonstrate the potential for recombinant scFv antibodies to be used as an alternative to the classical monoclonal antibodies or even molecular probes in the post-mortem analysis studies of generalist predators.

Monzo, Cesar; Urbaneja, Alberto; Ximenez-Embun, Miguel; Garcia-Fernandez, Julia; Garcia, Jose Luis; Castanera, Pedro

2012-01-01

378

Microheterogeneity of the collecting duct system in rabbit kidney as revealed by monoclonal antibodies  

Microsoft Academic Search

This report describes the immunolocalization of three monoclonal antibodies along the collecting duct system in rabbit kidney. The antibodies were raised against antigens derived from a membrane fraction of homogenized papillary tissue. Western Blot analysis demonstrated that each of the antibodies recognized a single band of about 190000 (PCD1), 210000 (PCD2) and 50000 (PCD3) daltons. In renal tissue, the antibodies

P. Gilbert; W. W. Minuth; S. Bachmann

1987-01-01

379

Removal of drugs from the circulation using immobilized monoclonal antibodies  

SciTech Connect

High-affinity monoclonal antidigoxin antibodies (dig-Ab) were immobilized to a pellicular microbead and characterized in terms of antibody affinity, specificity for other glycosides, and binding capacity. Determination of digoxin binding revealed that the binding capacity decreased to 25% of theoretical capacity. Attempts to improve the binding capacity were ineffective. A guinea pig animal model was developed to determine the efficacy of removing digoxin in vivo from the circulation using an antibody column. Male guinea pigs were hemoperfused with either a dig-Ab or bovine Y-globulin control column 16 h after a single i.v. injection of digoxin. Pre- and postcolumn plasma concentrations were obtained to evaluate the extraction efficiency. Hemoperfusion continued for 3 h at flow rates of 1.0-2.0 mL/min. Bound digoxin was eluted as described earlier and concentrations determined by (/sup 125/I) digoxin RIA. Amounts of digoxin removed represented less than 1% of the total body content. After several studies with the same column, the dig-Ab had lost most of its activity. A freshly prepared dig-Ab column removed approximately 20% of the total body content. Most of the measured constituents of the blood were unaffected by the procedure.

Brizgys, M.V.

1987-01-01

380

Modulation of catalysis and inhibition of fetal bovine serum acetylcholinesterase by monoclonal antibodies  

SciTech Connect

Monoclonal antibodies have been raised against acetyicholinesterase isolated from a variety of sources and species. Although none of these antibodies bind to the esteratic site, some of them appear to interact with the region of the catalytic subunit referred to as the peripheral anionic site. We describe here the production and characterization of six inhibitory monoclonal antibodies against fetal bovine serum acetylcholinesterase. Results show that changes in the conformation of acetyicholinesterase caused by interaction with monoclonal antibodies at a site remote from the catalytic site result in the modulation of catalytic activity of acetylcholinesterase.

Doctor, B.P.; Gentry, M.K.; Saxena, A.; Ashani, Y.

1995-12-31

381

Potential of palladium-109-labeled antimelanoma monoclonal antibody for tumor therapy  

SciTech Connect

Palladium-109, a beta-emitting radionuclide, was chelated to the monoclonal antibody 225.28S to the high molecular weight antigen associated with human melanoma. Injection of the radiolabeled monoclonal antibody into nude mice bearing human melanoma resulted in significant accumulation of the radiolabel in the tumors: 19% injected dose/g; 38:1 and 61:1 tumor-to-blood ratios at 24 and 48 hr, respectively. The localization of the radiolabeled antibody in liver and kidney also was high, but appreciably lower than that achieved in tumor. These results suggest Pd-109-labeled monoclonal antibody to tumor-associated antigens may have potential applications in tumor immunotherapy.

Fawwaz, R.A.; Wang, T.S.T.; Srivastava, S.C.; Rosen, J.M.; Ferrone, S.; Hardy, M.A.; Alderson, P.O.

1984-07-01

382

Advances in bispecific biotherapeutics for the treatment of cancer.  

PubMed

Conventional monoclonal antibody (mAb) therapeutics interfering with cellular signaling of their respective target antigens are frequently limited in their ability to induce significant anti-tumor activities when administered as single agents in patients with solid tumors. To overcome these limitations, several new technologies are being developed to empower biotherapeutics and to improve their anti-tumor activities, while maintaining their high tumor selectivity and superior safety profiles. The various efficacy enhancement technologies developed for mAbs can be divided broadly into two categories: First, technologies that improve the intrinsic anti-tumor activities of conventional immunoglobulin mAb formats, including the enhancement of effector cell functions and modulations of target binding properties, including interference with multiple signaling pathways. The second category of empowered biologics combines complementary anti-tumor modalities independent of the IgG format, including antibody drug conjugates (ADCs). In addition, bispecific compounds designed to recruit different subsets of inflammatory cells to the tumor environment, also belong to the mechanistic complementation strategy. This approach termed redirected immune cell killing, belongs to one the most promising new biotherapeutic platforms developed in oncology. Over 20 bispecific compounds are currently being developed pre-clinically, and several compounds are undergoing early stage clinical trials. In this report, we review the progress made in the development of bispecific biotherapeutics in the context of ADCs, redirected T- and B-cell killing and targeting of multiple signaling pathways. We also discuss the status of the clinical development of this class of compounds in oncology and the promises and challenges this field is currently facing. PMID:22858161

May, Chad; Sapra, Puja; Gerber, Hans-Peter

2012-11-01

383

Infectious complications associated with monoclonal antibodies and related small molecules.  

PubMed

Biologics are increasingly becoming part of routine disease management. As more agents are developed, the challenge of keeping track of indications and side effects is growing. While biologics represent a milestone in targeted and specific therapy, they are not without drawbacks, and the judicious use of these "magic bullets" is essential if their full potential is to be realized. Infectious complications in particular are not an uncommon side effect of therapy, whether as a direct consequence of the agent or because of the underlying disease process. With this in mind, we have reviewed and summarized the risks of infection and the infectious disease-related complications for all FDA-approved monoclonal antibodies and some related small molecules, and we discuss the probable mechanisms involved in immunosuppression as well as recommendations for prophylaxis and treatment of specific disease entities. PMID:19366915

Salvana, Edsel Maurice T; Salata, Robert A

2009-04-01

384

High-level iodination of monoclonal antibody fragments for radiotherapy  

SciTech Connect

Two different murine monoclonal antibody Fab fragments specific for p97, a melanoma-associated antigen, were labeled with I-131 at high activity levels without excessive chemical damage. Up to 20 mg of Fab were labeled with up to 300 mCi of I-131 using the chloramine-T method and large working volumes at room temperature. As much as 90% of the initial activity was recovered as labeled product. The labeled Fabs varied in their sensitivity to radioiodination damage, as measured by an in vitro cell-binding assay. Radioiodination was performed safely using a remote iodination apparatus. The final product was of radiopharmaceutical quality suitable for clinical diagnosis and experimental radiotherapy in humans.

Ferens, J.M.; Krohn, K.A.; Beaumier, P.L.; Brown, J.P.; Hellstroem, I.; Hellstroem, K.E.; Carrasquillo, J.A.; Larson, S.M.

1984-03-01

385

Positron emission tomographic imaging of tumors using monoclonal antibodies  

SciTech Connect

The overall objective for this research project is to develop methods for utilizing Positron Emission Tomography (PET) to increase the clinical potential of radiolabelled monoclonal antibodies (MAbs). By labeling MAbs with positron-emitting nuclides, it should be possible to quantitate the dynamics of their three-dimensional distribution in vivo. Our long term goals are to apply this approach to investigate the following: normal tissue toxicity; radiation dose to the tumor; and early tumor imaging. The research plans of this proposal include the following specific aims: optimize labeling of MAbs with fluorine 18, bromine 76 and bromine 75; label MAb Mel-14 (reactive against human gliomas and melanomas) and its Fab and F(ab{prime}){sub 2} fragments while retaining immunoreactivity; determine the distribution of Mel-14 in athymic mice bearing human gliomas; determine pharmacokinetics of Mel-14 in nonhuman primates. Experiments with another MAb, TP-1, and iodine 124 and 131 are also planned. 8 figs.

Zalutsky, M.R.

1990-12-01

386

Use of human monoclonal antibodies to treat Chikungunya virus infection.  

PubMed

Chikungunya virus (CHIKV) is an alphavirus prevalent in tropical regions. It causes an acute febrile disease that, in elderly individuals and newborns, is often associated with severe complications. We previously reported the isolation and characterization of 2 human monoclonal antibodies neutralizing CHIKV in vitro: 5F10 and 8B10. Here, we tested their efficacy in vivo as prophylactic and therapeutic treatments of CHIKV infection in AGR129 mice. In both settings, 5F10 and 8B10 were able to significantly delay CHIKV-driven lethality. Our results support the development of prophylactic and therapeutic treatments for CHIKV infection, using a combination of 5F10 and 8B10. PMID:23125446

Fric, Jan; Bertin-Maghit, Sébastien; Wang, Cheng-I; Nardin, Alessandra; Warter, Lucile

2013-01-15

387

Novel CD20 monoclonal antibodies for lymphoma therapy  

PubMed Central

Rituximab (RTX), a monoclonal antibody (mAb) against CD20, has been widely used for lymphoma therapy. RTX in combination with cyclophosphamide /doxorubicin /vincristine /prednisone (R-CHOP) remains the standard frontline regimen for diffuse large B-cell lymphoma. However, suboptimal response and /or resistance to rituximab have remained a challenge in the therapy of B-cell non-Hodgkin’s lymphoma (NHL). Novel agents are under active clinical trials. This review will summarize the latest development in new mAbs against CD20, which include second-generation mAbs, ofatumumab, veltuzumab (IMMU-106), ocrelizumab (PRO70769), and third-generation mAbs, AME-133v (ocaratuzumab), PRO131921 and GA101 (obinutumumab).

2012-01-01

388

Novel CD20 monoclonal antibodies for lymphoma therapy.  

PubMed

Rituximab (RTX), a monoclonal antibody (mAb) against CD20, has been widely used for lymphoma therapy. RTX in combination with cyclophosphamide /doxorubicin /vincristine /prednisone (R-CHOP) remains the standard frontline regimen for diffuse large B-cell lymphoma. However, suboptimal response and /or resistance to rituximab have remained a challenge in the therapy of B-cell non-Hodgkin's lymphoma (NHL). Novel agents are under active clinical trials. This review will summarize the latest development in new mAbs against CD20, which include second-generation mAbs, ofatumumab, veltuzumab (IMMU-106), ocrelizumab (PRO70769), and third-generation mAbs, AME-133v (ocaratuzumab), PRO131921 and GA101 (obinutumumab). PMID:23057966

Cang, Shundong; Mukhi, Nikhil; Wang, Kemeng; Liu, Delong

2012-01-01

389

Omalizumab: a recombinant humanized monoclonal IgE-blocking antibody.  

PubMed

Omalizumab (Xolair) is a recombinant humanized monoclonal antibody that blocks the high-affinity Fc receptor of immunoglobulin E (IgE). It is used to treat patients with moderate-persistent to severe-persistent asthma; patients must be older than 12 years, have a positive skin test to a perennial aeroallergen (e.g., dust mites, cats, dogs, and mold), and be symptomatic with inhaled corticosteroids. Omalizumab has a low incidence of side effects, and it is very expensive. The exact dosage of omalizumab dosage is determined by body weight and pretreatment serum total IgE levels. Omalizumab may have a role in the treatment of atopic dermatitis when IgE plays a causal role. PMID:15748543

Scheinfeld, Noah

2005-01-01

390

Monoclonal antibodies as a tool for phylogenetic studies of major histocompatibility antigens and beta 2-microglobulin.  

PubMed

The cross-reactivity of several monoclonal antibodies recognizing monomorphic determinants of human HLA-A, B, C, and DR antigens and human beta 2-microglobulin (beta 2m) has been studied on peripheral blood leukocytes in 24 different species. An monoclonal HLA-A-, B-, and C-specific antibody and four monoclonal HLA-DR-specific antibodies cross-reacted with cells from all the primate species tested. Furthermore, antibodies HLA-DR-specific were positive with peripheral blood leukocytes (PBL) from cows, goats, sheep, horses, and dogs. Two monoclonal beta 2m-specific antibodies, which were positive with PBL from certain primates, also reacted with cells from cows, goats, sheep, horses, and dogs. Two other beta 2-m-specific antibodies reacted only with PBL from chimpanzees. No reaction could be detected with all our reagents in other classes tested (birds, reptiles, amphibians, and Teleostei). PMID:6176537

Teillaud, J L; Crevat, D; Chardon, P; Kalil, J; Goujet-Zalc, C; Mahouy, G; Vaiman, M; Fellous, M; Pious, D

1982-01-01

391

Murine lupus monoclonal antibodies define five epitopes on two different Sm polypeptides.  

PubMed

Monoclonal anti-Sm (Smith) antibodies derived from the mouse strain MRL/lpr were isolated and characterized by binding to purified antigen, immunoprecipitating characteristic uridine-rich RNAs from Hela cell extracts, and by Western blot analysis using rabbit thymus extract. Five different Sm epitopes were demonstrated by epitope blockade and probing Western blots with the monoclonal antibodies. Human anti-Sm serum inhibited each monoclonal antibody from binding to antigen, indicating that both human and mouse antibodies bind to the same Sm epitopes. Human anti-Sm antibodies bound to 28,000 and 16,000 MW polypeptides, a small number also binding to a 14,000 MW polypeptide. The monoclonal antibodies also bound to the 28,000 and/or the 16,000 polypeptide, and provided evidence to suggest that these two Sm polypeptides bear some structural similarities, but are distinct molecules. PMID:2426187

Williams, D G; Stocks, M R; Smith, P R; Maini, R N

1986-07-01

392

Quality control of traditional chinese medicine by monoclonal antibody method.  

PubMed

In a previous study, we reported the preparation, characterization, variation, specificity, and sensitivity of an anti-aristolochic acid-II (AA-II) monoclonal antibody. The preparation procedure was as follows. AA-II conjugated with bovine serum albumin was used as an antigen for immunizing BALB/c mice. Splenocytes isolated from the immunized mice were fused with an aminopterin-sensitive mouse myeloma cell line to produce hybridoma cells secreting a mono-clonal antibody (MAb) against AA-II. The selected MAb was subsequently cloned. Hapten number, isotype, and an esti-mated dissociation constant (KD) of the secreted MAb were determined. This MAb was used to establish an ELISA method. The linear range was 0.19-13 µg/ml. Anti-AA-II MAb showed extremely high specificity for AA-II, low cross-reactivity (CR) against other AAs or aristololactam-I, and negligible CR (<0.5%) toward other natural compounds with different chemical structures. This study describes the successful application of the ELISA method using anti-AA-II MAb to determine AA-II concentration in several crude drugs derived from Aristolochia species. The highest AA-II concentration (2.82 µg/mg) was observed in the stem of A. manshuriensis, followed by that in the fruit of A. contorta (0.81 µg/mg). In case of A. indica, AA-II concentration in the root was higher than that in the aerial parts. These data indicated that the established ELISA method can be used for the quality control of crude drugs derived from Aristolochia plants. PMID:21143136

Shang, Ming-Ying; Tian, Min; Tanaka, Hiroyuki; Li, Xiao-Wei; Cai, Shao-Qing; Shoyama, Yukihiro

2011-03-01

393

Generation and characterization of monoclonal antibodies to equine NKp46  

PubMed Central

The immunoreceptor NKp46 is considered to be the most consistent marker of NK cells across mammalian species. Here, we use a recombinant NKp46 protein to generate a panel of monoclonal antibodies that recognize equine NKp46. The extracellular region of equine NKp46 was expressed with equine IL-4 as a recombinant fusion protein (rIL-4/NKp46) and used as an immunogen to generate mouse monoclonal antibodies (mAbs). MAbs were first screened by ELISA for an ability to recognize NKp46, but not IL-4, or the structurally related immunoreceptor CD16. Nine mAbs were selected and were shown to recognize full-length NKp46 expressed on the surface of transfected CHO cells as a GFP fusion protein. The mAbs recognized a population of lymphocytes by flow cytometric analysis that was morphologically similar to NKp46+ cells in humans and cattle. In a study using nine horses, representative mAb 4F2 labeled 0.8-2.1% PBL with a mean fluorescence intensity consistent with gene expression data. MAb 4F2+ PBL were enriched by magnetic cell sorting and were found to express higher levels of NKP46 mRNA than 4F2- cells by quantitative RT-PCR. CD3-depleted PBL from five horses contained a higher percentage of 4F2+ cells than unsorted PBL. Using ELISA, we determined that the nine mAbs recognize three different epitopes. These mAbs will be useful tools in better understanding the largely uncharacterized equine NK cell population.

Noronha, Leela E.; Harman, Rebecca M.; Wagner, Bettina; Antczak, Douglas F.

2012-01-01

394

Cellular determinants for preclinical activity of a novel CD33/CD3 bispecific T-cell engager (BiTE) antibody, AMG 330, against human AML.  

PubMed

CD33 is a valid target for acute myeloid leukemia (AML) but has proven challenging for antibody-drug conjugates. Herein, we investigated the cellular determinants for the activity of the novel CD33/CD3-directed bispecific T-cell engager antibody, AMG 330. In the presence of T cells, AMG 330 was highly active against human AML cell lines and primary AML cells in a dose- and effector to target cell ratio-dependent manner. Using cell lines engineered to express wild-type CD33 at increased levels, we found a quantitative relationship between AMG 330 cytotoxicity and CD33 expression; in contrast, AMG 330 cytotoxicity was neither affected by common CD33 single nucleotide polymorphisms nor expression of the adenosine triphosphate-binding cassette (ABC) transporter proteins, P-glycoprotein or breast cancer resistance protein. Unlike bivalent CD33 antibodies, AMG 330 did not reduce surface CD33 expression. The epigenetic modifier drugs, panobinostat and azacitidine, increased CD33 expression in some cell lines and augmented AMG 330-induced cytotoxicity. These findings demonstrate that AMG 330 has potent CD33-dependent cytolytic activity in vitro, which can be further enhanced with other clinically available therapeutics. As it neither modulates CD33 expression nor is affected by ABC transporter activity, AMG 330 is highly promising for clinical exploration as it may overcome some limitations of previous CD33-targeted agents. PMID:24311721

Laszlo, George S; Gudgeon, Chelsea J; Harrington, Kimberly H; Dell'Aringa, Justine; Newhall, Kathryn J; Means, Gary D; Sinclair, Angus M; Kischel, Roman; Frankel, Stanley R; Walter, Roland B

2014-01-23

395

Bispecific antibodies directed to CD4 domain 2 and HIV envelope exhibit exceptional breadth and picomolar potency against HIV-1  

PubMed Central

In the absence of an effective HIV-1 vaccine, passive immunization using broadly neutralizing Abs or Ab-like molecules could provide an alternative to the daily administration of oral antiretroviral agents that has recently shown promise as preexposure prophylaxis. Currently, no single broadly neutralizing Ab (bNAb) or combination of bNAbs neutralizes all HIV-1 strains at practically achievable concentrations in vivo. To address this problem, we created bispecific Abs that combine the HIV-1 inhibitory activity of ibalizumab (iMab), a humanized mAb directed to domain 2 of human CD4, with that of anti-gp120 bNAbs. These bispecific bNAbs (BibNAbs) exploit iMab’s potent anti–HIV-1 activity and demonstrated clinical efficacy and safety to anchor and thereby concentrate a second broadly neutralizing agent at the site of viral entry. Two BibNabs, PG9-iMab and PG16-iMab, exhibit exceptional breadth and potency, neutralizing 100% of the 118 viruses tested at low picomolar concentrations, including viruses resistant to both parental mAbs. The enhanced potency of these BibNAbs was entirely dependent on CD4 anchoring, not on membrane anchoring per se, and required optimal Ab geometry and linker length. We propose that iMab-based BibNAbs, such as PG9-iMab and PG16-iMab, are promising candidates for passive immunization to prevent HIV-1 infection.

Pace, Craig S.; Song, Ruijiang; Ochsenbauer, Christina; Andrews, Chasity D.; Franco, David; Yu, Jian; Oren, Deena A.; Seaman, Michael S.; Ho, David D.

2013-01-01

396

Impact of Polymorphisms on the Clinical Outcomes of Monoclonal Antibody Therapy Against Hematologic Malignancies  

Microsoft Academic Search

\\u000a Monoclonal antibodies that target various specific antigens can be used to kill the tumor cells expressing specific antigens,\\u000a especially in hematologic cancers. Rituximab, one of the commonly used monoclonal antibodies, was suggested to mediate its\\u000a action mechanism via antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), or a direct\\u000a pro-apoptotic effect. It has been proposed that the inter-individual variation of gene

Dong Hwan Kim

397

Monoclonal antibodies against a component related to soluble estrogen receptor.  

PubMed

Mice were immunized with 1 to 3 micrograms of cytoplasmic estrogen receptor fragment purified from human myometrium by affinity chromatography. Two RE-antibody-secreting clones were detected from one fusion that were capable of precipitating cytosol RE. Monoclonal antibody D5 (subclass IgG1) reacts with an antigen that is related to RE from immunoprecipitation studies but which can be separated from the hormone binding unit. In the presence of anti-mouse serum, D5 precipitates labeled human cytoplasmic RE complexes from breast tumor, fibroid, myometrial, and endometrial preparations but does not react with nuclear RE from human endometrium or cytoplasmic RE from other species tested. Conversely, antibody C3 (Class IgM) precipitates human cytoplasmic RE and nuclear RE complexes as well as labeled cytoplasmic RE from rat and calf uterus and chick oviduct. Neither antibody reacts with progesterone receptor or androgen receptor from human breast tumor, SHBG from human plasma, or rat alpha-fetoprotein. With D5, steroid labeling of cytoplasmic RE at 25 degrees increased the RE immune complex precipitated. D5 precipitates molybdate-stabilized RE from myometrial cytosol when labeled at 25 degrees but not at 4 degrees. C5 precipitates molybdate-stabilized RE whether cytosol was steroid-labeled at 4 degrees or 25 degrees. For D5, optimal precipitation of RE from human breast tumor was observed when cytosol was steroid-labeled at 25 degrees in buffers of pH range 5 to 6. Immunochemical studies indicate that D5 is associated with a Mr 29,000 component in RE-positive cytosols. Electrofocusing and sucrose density gradient analysis confirmed that D5 antigen is a non-hormone-binding component related to cytosolic RE from breast tumor and myometrium. PMID:4016746

Coffer, A I; Lewis, K M; Brockas, A J; King, R J

1985-08-01

398

Metastatic colorectal cancer cells from patients previously treated with chemotherapy are sensitive to T-cell killing mediated by CEA\\/CD3-bispecific T-cell-engaging BiTE antibody  

Microsoft Academic Search

Background:Novel technologies to redirect T-cell killing against cancer cells are emerging. We hypothesised that metastatic human colorectal cancer (CRC) previously treated with conventional chemotherapy would be sensitive to T-cell killing mediated by carcinoembryonic antigen (CEA)\\/CD3-bispecific T-cell-engaging BiTE antibody (MEDI-565).Methods:We analysed proliferation and lysis of CEA-positive (CEA+) CRC specimens that had survived previous systemic chemotherapy and biologic therapy to determine whether

T Osada; D Hsu; S Hammond; A Hobeika; G Devi; T M Clay; H K Lyerly; M A Morse

2010-01-01

399

Combination of rituximab with blinatumomab (MT103\\/MEDI538), a T cell-engaging CD19-\\/CD3-bispecific antibody, for highly efficient lysis of human B lymphoma cells  

Microsoft Academic Search

We have compared the cytotoxic activity of rituximab with that of blinatumomab (MT103\\/MEDI-538), a single-chain CD19-\\/CD3-bispecific antibody engaging human T cells. Blinatumomab consistently led to a higher degree of lysis of human lymphoma lines than rituximab, and was active at much lower concentration. The cytotoxicity mediated by blinatumomab and rituximab both caused a potent activation of pro-caspases 3 and 7

Sandrine d’Argouges; Sandra Wissing; Christian Brandl; Nadja Prang; Ralf Lutterbuese; Alex Kozhich; JoAnn Suzich; Mathias Locher; Peter Kiener; Peter Kufer; Robert Hofmeister; Patrick A. Baeuerle; Ralf C. Bargou

2009-01-01

400

Anti-idiotypic responses of lactating cows immunized with monoclonal antibodies against bovine somatotropin.  

PubMed

The objective of this study was to produce anti-idiotypic antibodies with bovine somatotropin (bST)-like activity by active immunization of lactating cows and to determine their effects on milk yield. Several monoclonal antibodies against bST were evaluated for their interaction with bST in a rat growth bioassay. Two bST-agonist monoclonal antibodies (1 and 2), and two bST-antagonist monoclonal antibodies (3 and 4) were selected. Cows were immunized with immunoglobulin G as a control (n = 12) or with one of the four anti-bST monoclonal antibodies (1, 2, 3, 4; n = 12) on d 3, 24, 45, 66, 87, 108, 129, and 150 of lactation. From wk 3 of lactation, all cows immunized with each of the four anti-bST monoclonal antibodies developed anti-idiotypes until wk 30 of lactation. Total lactation yields were not different among monoclonal antibodies 2, 3, and 4 and control cows (9299, 9321, 9733, and 9415 kg, respectively). However, cows immunized with anti-bST monoclonal antibody 1 had reduced lactation yield compared with cows on other treatments (8136 kg). Daily milk yield of cows immunized with monoclonal antibody 1 was decreased from wk 9 of lactation [36.2 vs. 40.9 kg/d (control)] until the end of lactation, concomitantly with decreased bST concentration from wk 9 of lactation. Cows immunized with anti-bST monoclonal antibody 4 had increased milk yield compared with that of controls during wk 3 to 6 and wk 18 to 21 of lactation. Therefore, anti-idiotypes directed against anti-bST 1 had bST-antagonistic effects on lactation performance; anti-idiotypes against anti-bST 4 transiently increased milk yield. PMID:10480096

Roberge, S; Wilkie, B N; Walton, J S

1999-08-01

401

Antibody-guided irradiation of advanced ovarian cancer with intraperitoneal monoclonal antibodies  

SciTech Connect

Twenty-four patients with persistent epithelial ovarian cancer after chemotherapy with or without external beam irradiation, were treated with intraperitoneally administered /sup 131/I-labeled monoclonal antibodies HMFG1, HMFG2, AUA1, H17E2, directed against tumor-associated antigens. Acute side effects were mild abdominal pain, pyrexia, diarrhea, and moderate reversible pancytopenia. One patient developed a subphrenic abscess requiring surgical drainage. Eight patients with large volume disease, ie, greater than 2 cm tumor diameter, did not respond to antibody-guided irradiation and died of progressive disease within 9 months of treatment. Sixteen patients had small-volume (less than 2 cm) disease at the time of treatment with radiolabeled antibody. Seven patients failed to respond, and of nine initial responders, four patients remain alive and free from disease 6 months to 3 years from treatment. Analysis of the data on relapse indicated that doses greater than 140 mCi were more effective than lower doses. We conclude that the intraperitoneal administration of 140 mCi or more of /sup 131/I-labeled tumor-associated monoclonal antibodies represents a new and potentially effective form of therapy for patients with small-volume stage III ovarian cancer.

Epenetos, A.A.; Munro, A.J.; Stewart, S.; Rampling, R.; Lambert, H.E.; McKenzie, C.G.; Soutter, P.; Rahemtulla, A.; Hooker, G.; Sivolapenko, G.B.

1987-12-01

402

Influence of unlabeled monoclonal anti-mouse antibody on the clearance rate of radiolabeled mouse monoclonal antibody  

SciTech Connect

High blood background levels of intact radiolabeled monoclonal antibody (MoAb) after intravenous (iv) injection are problematic. The injection of unlabeled polyclonal antimouse Abs following injection with labeled MoAbs produces accelerated MoAb clearance. This study evaluates a Mo antimouse Ab for efficacy of accelerating radio MoAb clearance. HB-58 is a rat/mouse MoAb which binds strongly to mouse kappa light chains present in 95% of murine monoclonals. It is unreactive with rat, rabbit or human kappa chains. Six rats were injected iv with 30 ..mu..Ci (approximately 6 ..mu..g) of I-125 UPC-10, a non-specific IgG2ak MoAb that is bound to well by HB-58. No alteration was seen in the clearance of UPC-10 in any of the animals, regardless of the injection type or amount on the second day. In addition, no increase in liver or spleen activity was seen in those rats that received HB-58. The lack of change in rate of clearance and biodistribution of UPC-10 after the iv injection of a purified, specific, anti-mouse MoAb is in marked contrast to the accelerated clearance reported following polyclonal anti-mouse antibody administration. This may be due to the inability of MoAbs to cross link. These preliminary studies suggest that Mo anti-mouse Abs, at these dose levels, are not useful in achieving increased rates of radiolabeled murine MoAb clearance.

Wahl, R.L.; Laino, L.; Jackson, G.; Fisher, S.; Beierwaltes, W.H.

1985-05-01

403

Monoclonal antibody (M2) to glial and neuronal cell surfaces.  

PubMed

A monoclonal antibody designated M2 arose from the fusion of mouse myeloma cells with splenocytes from a rat immunized with particulate fraction from early postnatal mouse cerebellum. Expression of M2 antigen was examined by indirect immunofluorescence on frozen sections of developing and adult mouse cerebellum and on monolayer cultures of early postnatal mouse cerebellar cells. In adult cerebellum, M2 staining outlines the cell bodies of granule and Purkinje cells. A weaker, more diffuse staining is seen in the molecular layer and white matter. In sections of newborn cerebellum, M2 antigen is weakly detectable surrounding cells of the external granular layer and Purkinje cells. The expression of M2 antigen increases during development in both cell types, reaching adult levels by postnatal day 14. At all stages of postnatal cerebellar development, granule cells that have completed migration to the internal granule layer are more heavily stained by M2 antibodies than are those before and in process of migration. In monolayer cultures, M2 antigen is detected on the cell surface of all GFA protein-positive astrocytes and on more immature oligodendrocytes that express 04 antigen but not 01 antigen. After 3 days in culture, tetanus toxin positive neurons begin to express M2 antigen. The same delayed expression of M2 antigen on neurons is observed in cultures derived from mice ranging in age from postnatal day 0 to 10. PMID:6170757

Lagenaur, C; Schachner, M

1981-01-01

404

Defining process design space for monoclonal antibody cell culture.  

PubMed

The concept of design space has been taking root as a foundation of in-process control strategies for biopharmaceutical manufacturing processes. During mapping of the process design space, the multidimensional combination of operational variables is studied to quantify the impact on process performance in terms of productivity and product quality. An efficient methodology to map the design space for a monoclonal antibody cell culture process is described. A failure modes and effects analysis (FMEA) was used as the basis for the process characterization exercise. This was followed by an integrated study of the inoculum stage of the process which includes progressive shake flask and seed bioreactor steps. The operating conditions for the seed bioreactor were studied in an integrated fashion with the production bioreactor using a two stage design of experiments (DOE) methodology to enable optimization of operating conditions. A two level Resolution IV design was followed by a central composite design (CCD). These experiments enabled identification of the edge of failure and classification of the operational parameters as non-key, key or critical. In addition, the models generated from the data provide further insight into balancing productivity of the cell culture process with product quality considerations. Finally, process and product-related impurity clearance was evaluated by studies linking the upstream process with downstream purification. Production bioreactor parameters that directly influence antibody charge variants and glycosylation in CHO systems were identified. PMID:20589669

Abu-Absi, Susan Fugett; Yang, LiYing; Thompson, Patrick; Jiang, Canping; Kandula, Sunitha; Schilling, Bernhard; Shukla, Abhinav A

2010-08-15

405

Evidence of a saturable hepatic receptor for mouse monoclonal antibodies  

SciTech Connect

Monoclonal antibodies (MAb) can be labeled with I-123 at high specific activities, so that large amounts of radioactivity attached to small amounts of protein can be injected for radioimmunoimaging. This conserves antibody and decreases the opportunity for foreign protein reactions and target tissue binding site saturation. In order to assess the effects on pharmacokinetics and imaging, the authors administered microgram amounts of I-123-MAb (Lyn-1, IgG2a or B6.01, IgGl with and following 4-5 milligram preloading with MAb on separate occasions to 4 patients with a target tumor (B cell lymphoma) and 2 patients without a target tumor (breast cancer). Pharmacokinetics were observed in blood and urine by counting whole samples and HPLC fractions of these samples and in organs by serial imaging. Early blood clearance and urinary excretion were faster after injection of microgram amounts of MAb, but subsequently were comparable to those obtained after preload. This paper concludes that the amount of administered MAb dramatically influences the pharmacokinetics of mouse MAb. Saturable hepatic Fc receptors are probably the source of these observations. Reports of accelerated deiodination of MAb are related to this phenomenon. Optimal imaging and treatment with MAb requires saturation of these hepatic receptors.

De Nardo, G.L.; De Nardo, S.J.; Peng, J.S.; O'Grady, L.F.; Mills, S.L.; Epstein, A.L.; Cardiff, R.D.

1985-05-01

406

Generation and characterization of monoclonal antibodies to equine CD16  

PubMed Central

The low-affinity Fc receptor CD16 plays a central role in the inflammatory and innate immune responses of many species, but has not yet been investigated in the horse. Using the predicted extracellular region of equine CD16 expressed as a recombinant fusion protein with equine IL-4 (rIL-4/CD16), we generated a panel of mouse monoclonal antibodies (mAbs) that recognize equine CD16. Nine mAbs were chosen for characterization based upon recognition of CD16, but not IL-4, in ELISA. All nine mAbs recognized full-length, cell-surface CD16 expressed as a GFP fusion protein by CHO cells, but not the closely related Fc receptor CD32 expressed in the same system. In flow cytometric analysis with equine peripheral leukocytes, the mAbs labeled cells in the granulocyte, monocyte, and lymphocyte populations in a pattern consistent with other species. Monocytes that were strongly labeled with CD16 mAb 9G5 were also positive for the LPS receptor CD14. Cytospins made with peripheral leukocytes were immunohistochemically labeled and showed mAb recognition of primarily mononuclear cells. ELISA revealed that the nine mAbs can be grouped into three patterns of epitope recognition. These new antibodies will serve as useful tools in the investigation of equine immune responses and inflammatory processes.

Noronha, Leela E.; Harman, Rebecca M.; Wagner, Bettina; Antczak, Douglas F.

2012-01-01

407

Monoclonal antibody-based therapies for microbial diseases  

PubMed Central

The monoclonal antibody (mAb) revolution that currently provides many new options for the treatment of neoplastic and inflammatory diseases has largely bypassed the field of infectious diseases. Only one mAb is licensed for use against an infectious disease, although there are many in various stages of development. This situation is peculiar given that serum therapy was one of the first effective treatments for microbial diseases and that specific antibodies have numerous antimicrobial properties. The underdevelopment and underutilization of mAb therapies for microbial diseases has various complex explanations that include the current availability of antimicrobial drugs, small markets, high costs and microbial antigenic variation. However, there are signs that the climate for mAb therapeutics in infectious diseases is changing given increasing antibiotic drug resistance, the emergence of new pathogenic microbes for which no therapy is available, and development of mAb cocktail formulations. Currently, the major hurdle for the widespread introduction of mAb therapies for microbial diseases is economic, given the high costs of immunoglobulin preparations and relatively small markets. Despite these obstacles there are numerous opportunities for mAb development against microbial diseases and the development of radioimmunotherapy provides new options for enhancing the magic bullet. Hence, there is cautious optimism that the years ahead will see more mAbs in clinical use against microbial diseases.

Saylor, Carolyn; Dadachova, Ekaterina; Casadevall, Arturo

2009-01-01

408

Production of human anti-HLA monoclonal antibodies  

SciTech Connect

Only 40% of the several hundred anti-HLA murine monoclonal antibodies (MAbs) that have been made detect HLA-A,B,C or DR specificities previously defined by human alloantisera, the range of recognized specificities is very narrow, and few of the MAbs have proven useful as tissue typing reagents. In hopes of obtaining HLA typing reagents, the authors are developing a protocol for the production of human anti-HLA MAbs from HLA-antigen (Ag) immunized peripheral blood B cells of volunteering renal patients, immunized to one or more HLA Ags through therapeutic blood transfusions. A simple enrichment of the donor B cells has not been sufficient for anti-HLA MAb production, the authors are currently delineating the conditions necessary for increasing the number of HLA-specific donor B cells by in vitro stimulation with cells expressing the HLA Ag to which the B cell donor is immunized. For the production of MAbs, the stimulated B cells are transformed with Epstein-Barr virus and subsequently fused with KR-4 lymphoblastoid cells. Hybridomas are selected by HAT and Ouabain. Supernatants are screened for anti-HLA activity against lymphocyte targets expressing the original immunizing HLA Ag by complement mediated /sup 51/Cr release assay. Antibody specificity is determined by the complement-dependent microcytotoxicity test used for HLA typing.

Walker, M.C.; Mercier, F.; Roger, J.; Varin, M.

1986-03-01

409

Radiolabeled monoclonal antibodies for imaging and therapy: Potential, problems, and prospects: Scientific highlights  

SciTech Connect

This meeting focused on areas of research on radiolabeled monoclonal antibodies. Topics covered included the production, purification, and fragmentation of monoclonal antibodies and immunochemistry of hybridomas; the production and the chemistry of radionuclides; the radiohalogenation and radiometal labeling techniques; the in-vivo pharmacokinetics of radiolabeled antibodies; the considerations of immunoreactivity of radiolabeled preparations; the instrumentation and imaging techniques as applied to radioimmunodetection; the radiation dosimetry in diagnostic and therapeutic use of labeled antibodies; the radioimmunoscintigraphy and radioimmunotherapy studies; and perspectives and directions for future research. Tutorial as well as scientific lectures describing the latest research data on the above topics were presented. Three workshop panels were convened on ''Methods for Determining Immunoreactivity of Radiolabeled Monoclonal Antibodies - Problems and Pitfalls,'' Radiobiological and Dosimetric Considerations for Immunotherapy with Labeled Antibodies,'' and ''The Human Anti-Mouse Antibody Response in Patients.''

Srivastava, S.C.; Buraggi, G.L.

1986-01-01

410

Structure of solid tumors and their vasculature: Implications for therapy with monoclonal antibodies  

SciTech Connect

Delivery of monoclonal antibodies to solid tumors is a vexing problem that must be solved if these antibodies are to realize their promise in therapy. Such success as has been achieved with monoclonal antibodies is attributable to the local hyperpermeability of the tumor vasculature, a property that favors antibody extravasation at tumor sites and that is mediated by a tumor-secreted vascular permeability factor. However, leaky tumor blood vessels are generally some distance removed from target tumor cells, separated by stroma and by other tumor cells that together represent significant barriers to