학술논문

Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens
Document Type
Academic Journal
Source
Proceedings of the National Academy of Sciences of the United States. May 26, 1998, Vol. 95 Issue 11, p6157, 6 p.
Subject
Bacteriophages -- Analysis
Antigen-antibody reactions -- Research
Antigens -- Research
Science and technology
Language
ISSN
0027-8424
Abstract
A large library of phage-displayed human single-chain Fv antibodies (scFv), containing 6.7 x [10.sup.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.