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 1046-2023Full text not available from this repository.
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 ? expression vectors. The libraries of ? Fd and ? light chains are then combined to generate a single ? combinatorial vector encoding both chains and capable of generating a Fab fragment. Since the libraries are constructed in bacteriophage ?, 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.
|Subjects:||University of Westminster > Science and Technology > Life Sciences, School of (No longer in use)|
|Depositing User:||Miss Nina Watts|
|Date Deposited:||07 Jan 2008|
|Last Modified:||23 Dec 2009 14:27|
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