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Chimeric antibody with specificity to human B cell surface antigen

a chimeric antibody and b cell surface technology, applied in the field of recombinant dna, can solve the problems of harmful hypersensitivity reactions, increased clearance rate of circulating antibody molecules, and many obstacles to the development of human monoclonal antibodies

Inactive Publication Date: 2005-07-28
ROBINSON RANDY +2
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

Unfortunately, a number of obstacles exist with respect to the development of human monoclonal antibodies.
Although such antibodies may be used therapeutically in man, repeated injections of “foreign” antibodies, such as a mouse antibody, in humans, can lead to harmful hypersensitivity reactions as well as increased rate of clearance of the circulating antibody molecules so that the antibodies do not reach their target site.
Furthermore, mouse monoclonal antibodies may lack the ability to efficiently interact with human effector cells as assessed by functional assays such as antibody-dependent cellular cytotoxicity (ADCC) and complement-mediated cytolysis (CDC).
Another problem faced by immunologists is that most human monoclonal antibodies obtained in cell culture are of the IgM type.
However, an unexpected splice donor site caused the deletion of the V region leader sequence.
Thus, this approach did not yield complete chimeric antibody molecules.

Method used

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  • Chimeric antibody with specificity to human B cell surface antigen
  • Chimeric antibody with specificity to human B cell surface antigen
  • Chimeric antibody with specificity to human B cell surface antigen

Examples

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example 1

A Chimeric Mouse-Human Immunoglobulin with Specificity for a Human B-Cell Surface Antigen

[0137] (1) Antibody 2H7.

[0138] The 2H7 mouse monoclonal antibody (gamma 2b, kappa) recognizes a human B-cell surface antigen, (Bp35(CD20)) Clark, E. A., et al., Proc. Natl. Acad. Sci., U.S.A. 82: 1766 (1985)). The (Bp35(CD20)) molecules presumably play a role in B-cell activation. The antibody 2H7 does not react with stem cells which are progenitors of B-cells epithelial, mesenchymal and fibroblastic cells of other organs.

[0139] (2) Identification of J Sequences in the Immunoglobulin mRNA of 2H7.

[0140] Frozen cells were thawed on ice for 10 minutes and then at room temperature. The suspension was diluted with 15 ml PBS and the cells were centrifuged down. They were resuspended, after washes in PBS, in 16 ml 3M LiCl, 6M urea and disrupted in a polytron shear. The preparation of mRNA and the selection of the poly(A+) fraction were carried out according to Auffray, C. and Rougeon, F., Eur. J. B...

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Abstract

A chimeric antibody with human constant region and murine variable region, having specificity to a 35 kDA polypeptide (Bp35(CD20)) expressed on the surface of human B cells, methods of production, and uses.

Description

[0001] This application is a continuation in part of U.S. application Ser. No. ______, International Application No. PCT / US86 / 02269, filed Oct. 27, 1986, in the PCT receiving office of the U.S.BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] This invention relates to recombinant DNA methods of preparing an antibody with specificity for an antigen on the surface of human B cells, genetic sequences coding therefor, as well as methods of obtaining such sequences. [0004] 2. Background Art [0005] The application of cell-to-cell fusion for the production of monoclonal antibodies by Kohler and Milstein (Nature (London), 256: 495, 1975) spawned a revolution in biology equal in impact to that from the invention of recombinant DNA cloning. Monoclonal antibodies produced from hybridomas are already widely used in clinical and basic scientific studies. Applications of human monoclonal antibodies produced by human hybridomas hold great promise for the treatment of cancer, vira...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00A61K47/48C07K16/28C07K16/46
CPCA61K38/00A61K47/48561A61K47/48669A61K2039/505C07K16/2887C07K16/462C07K2319/40C07K2317/732C07K2317/734C07K2319/00C07K2319/02C07K2319/30C07K2317/24A61K47/6849A61K47/6877
Inventor ROBINSON, RANDYLIU, ALVINLEDBETTER, JEFFREY
Owner ROBINSON RANDY
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