NDLI: A binary plasmid system for shuffling combinatorial antibody libraries.

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On the nature of global classification.

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A binary plasmid system for shuffling combinatorial antibody libraries.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	O'kennedy, R. Roben, P. Barbas, C. F. Lerner, R. A. Collet, T. A. Burton, D. R.
Description	Author Affiliation: Collet TA ( Department of Chemistry, Scripps Research Institute, La Jolla, CA 92037.);
Abstract	We have used a binary system of replicon-compatible plasmids to test the potential for promiscuous recombination of heavy and light chains within sets of human Fab fragments isolated from combinatorial antibody libraries. Antibody molecules showed a surprising amount of promiscuity in that a particular heavy chain could recombine with multiple light chains with retention of binding to a protein antigen. The degree to which a given heavy chain productively paired with any light chain to bind antigen varied from 43% to 100% and depended strongly on the heavy-chain sequence. Such productive crosses resulted in a set of Fab fragments of similar apparent binding constants, which seemed to differ mainly in the amount of active Fab fragment produced in the bacterial cell. The dominance of the heavy chain in the antibody-antigen interaction was further explored in a set of directed crosses, in which heavy and light chains derived from antigen-specific clones were crossed with nonrelated heavy and light chains. In these crosses, an Fab fragment retained antigen binding only if it contained a heavy chain from an antigen-specific clone. In no case did the light chain confer detectable affinity when paired with indifferent heavy chains. The surprising promiscuity of heavy chains has ramifications for the evaluation of the diversity of combinatorial libraries made against protein antigens and should allow the combination of one such promiscuous heavy chain with an engineered light chain to form an Fab fragment carrying synthetic cofactors to assist in antibody catalysis.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	21
Volume Number	89
Language	English
Publisher	National Academy of Sciences
Publisher Date	1992-12-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Antibodies Genetics Gene Library Immunoglobulin Fab Fragments Immunoglobulin Heavy Chains Immunoglobulin Light Chains Plasmids Amino Acid Sequence Enzyme-Linked Immunosorbent Assay Gene Products, Env Immunology HIV HIV Envelope Protein Gp120 HIV Envelope Protein Gp160 Molecular Sequence Data Oligodeoxyribonucleotides Promoter Regions, Genetic Protein Precursors Restriction Mapping Research Support, Non-U.S. Gov't Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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