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

Nat Biotechnol. 2007 May;25(5):563-5. doi: 10.1038/nbt1296. Epub 2007 Apr 15.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / isolation & purification*
  • Antibodies, Monoclonal / physiology*
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Humans
  • Peptide Library*
  • Protein Engineering / methods*

Substances

  • Antibodies, Monoclonal
  • Peptide Library