Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies

Construction of a heavy chain variable region-specific cDNA library

L. Sastry, M. Alting-Mees, W. D. Huse, J. M. Short, J. A. Sorge, B. N. Hay, K. D. Janda, Stephen Benkovic, R. A. Lerner

Research output: Contribution to journalArticle

160 Citations (Scopus)

Abstract

Efficient generation of catalytic antibodies is uniquely dependent on the exact nature of the binding interactions in the antigen-antibody complex. Current methods for generation of monoclonal antibodies do not efficiently survey the immunological repertoire and, therefore, they limit the number of catalysts that can be obtained. We are exploring methods to clone and express the immunological repertoire in Escherichia coli. As the essential first step, we present here a method for the establishment of a highly diverse heavy chain variable region library. Consequently, it should now be possible to express and recombine the heavy and light chain variable region fragments to generate a large array of functional combining portions of the antibody molecule. This technology may provide an alternative to the hybridoma methodology for accessing the monoclonal antibody specificity of the immune system.

Original languageEnglish (US)
Pages (from-to)5728-5732
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number15
DOIs
StatePublished - Jan 1 1989

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Catalytic Antibodies
Gene Library
Organism Cloning
Monoclonal Antibodies
Escherichia coli
Antibody Specificity
Hybridomas
Antigen-Antibody Complex
Libraries
Immune System
Clone Cells
Technology
Light
Antibodies

All Science Journal Classification (ASJC) codes

  • General

Cite this

Sastry, L. ; Alting-Mees, M. ; Huse, W. D. ; Short, J. M. ; Sorge, J. A. ; Hay, B. N. ; Janda, K. D. ; Benkovic, Stephen ; Lerner, R. A. / Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies : Construction of a heavy chain variable region-specific cDNA library. In: Proceedings of the National Academy of Sciences of the United States of America. 1989 ; Vol. 86, No. 15. pp. 5728-5732.
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Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies : Construction of a heavy chain variable region-specific cDNA library. / Sastry, L.; Alting-Mees, M.; Huse, W. D.; Short, J. M.; Sorge, J. A.; Hay, B. N.; Janda, K. D.; Benkovic, Stephen; Lerner, R. A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 86, No. 15, 01.01.1989, p. 5728-5732.

Research output: Contribution to journalArticle

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T1 - Cloning of the immunological repertoire in Escherichia coli for generation of monoclonal catalytic antibodies

T2 - Construction of a heavy chain variable region-specific cDNA library

AU - Sastry, L.

AU - Alting-Mees, M.

AU - Huse, W. D.

AU - Short, J. M.

AU - Sorge, J. A.

AU - Hay, B. N.

AU - Janda, K. D.

AU - Benkovic, Stephen

AU - Lerner, R. A.

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