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Combinatorial immunoglobulin libraries in phage

Kang, Angray S. and Burton, Dennis R. and Lerner, Richard A. (1991) Combinatorial immunoglobulin libraries in phage. Methods: a Companion to Methods in Enzymology, 2 (2). pp. 111-118. ISSN 1046-2023

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Official URL: http://dx.doi.org/10.1016/S1046-2023(05)80211-7

Abstract

A rapid route for the generation of monoclonal antibodies by repertoire cloning is described. The technique circumvents the need for conventional hybridoma production and allows a variety of new approaches in antibody design and selection. The genes encoding the Fd and light chains are accessed by PCR amplification from cDNA and cloned into two separate λ expression vectors. The libraries of λ Fd and λ light chains are then combined to generate a single λ combinatorial vector encoding both chains and capable of generating a Fab fragment. Since the libraries are constructed in bacteriophage λ, in vitro packaging offers a very efficient route for reintroducing the recombinant DNA back into Escherichia coli. The library generated in such a way may be screened by conventional plaque lifts on nitrocellulose filters using labeled antigen.

Item Type:Article
Research Community:University of Westminster > Life Sciences, School of
ID Code:4544
Deposited On:07 Jan 2008
Last Modified:23 Dec 2009 14:27

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