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PMID: 9600934 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 11 ·1998-05-26 ·Pages 6157-62

Sheets MD, Amersdorfer P, Finnern R, Sargent P, Lindquist E, Schier R, Hemingsen G, Wong C, Gerhart JC, Marks JD, Lindqvist E

Abstract

A large library of phage-displayed human single-chain Fv antibodies (scFv), containing 6.7 x 10(9) members, was generated by improving the steps of library construction. Fourteen different protein antigens were used to affinity select antibodies from this library. A panel of specific antibodies was isolated with each antigen, and each panel contained an average of 8.7 different scFv. Measurements of antibody-antigen interactions revealed several affinities below 1 nM, comparable to affinities observed during the secondary murine immune response. In particular, four different scFv recognizing the ErbB2 protein had affinities ranging from 220 pM to 4 nM. Antibodies derived from the library proved to be useful reagents for immunoassays. For example, antibodies generated to the Chlamydia trachomatis elementary bodies stained Chlamydia-infected cells, but not uninfected cells. These results demonstrate that phage antibody libraries are ideally suited for the rapid production of panels of high-affinity mAbs to a wide variety of protein antigens. Such libraries should prove especially useful for generating reagents to study the function of gene products identified by genome projects.

MeSH Terms
Animals Antibodies/genetics,immunology Antibody Affinity Antigens/immunology Gene Library Humans Immunoglobulin Variable Region/genetics,immunology Mice Proteins/immunology
Chemicals
Antibodies Antigens Immunoglobulin Variable Region Proteins
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Sheets M D
Department of Molecular Cell Biology, University of California, Berkeley, CA 94720, USA.
Amersdorfer P
Finnern R
Sargent P
Lindquist E
Schier R
Hemingsen G
Wong C
Gerhart J C
Marks J D
Lindqvist E
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.11.6157
Published
1998-05-26
Pages
6157-62
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC27609
Subset
IM
Grants
NIGMS NIH HHS · GM15203-02 · United States
NIGMS NIH HHS · R01 GM19363 · United States
Corrections
ErratumIn
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External Links
PubMed source DOI: 10.1073/pnas.95.11.6157 PMC Full Text View journal details
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