Title | Combinatorial immunoglobulin libraries in phage |
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Authors | Kang, A.S., Burton, D.R. and Lerner, R.A. |
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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. |
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Journal | Methods: a Companion to Methods in Enzymology |
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Journal citation | 2 (2), pp. 111-118 |
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ISSN | 1046-2023 |
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Year | Apr 1991 |
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Digital Object Identifier (DOI) | https://doi.org/10.1016/S1046-2023(05)80211-7 |
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Publication dates |
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Published | Apr 1991 |
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