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Nat Biotechnol. 2007 May;25(5):563-5. Epub 2007 Apr 15.

Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli.

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1
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.

Abstract

We describe facile isolation of full-length IgG antibodies from combinatorial libraries expressed in E. coli. Full-length heavy and light chains are secreted into the periplasm, where they assemble into aglycosylated IgGs that are captured by an Fc-binding protein that is tethered to the inner membrane. After permeabilizing the outer membrane, spheroplast clones expressing so-called E-clonal antibodies, which specifically recognize fluorescently labeled antigen, are selected using flow cytometry. Screening of a library constructed from an immunized animal yielded several antibodies with nanomolar affinities toward the protective antigen of Bacillus anthracis.

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PMID:
17435747
DOI:
10.1038/nbt1296
[Indexed for MEDLINE]

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