Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli.
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.
Mazor
Yariv
Y
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
Van Blarcom
Thomas
T
Mabry
Robert
R
Iverson
Brent L
BL
Georgiou
George
G
D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods*
D000911:Antibodies, Monoclonal; D019151:Peptide Library