Efficient construction of a large nonimmune phage antibody library: The production of high-affinity human single-chain antibodies to protein antigens
| Autor: | M D, Sheets, P, Amersdorfer, R, Finnern, P, Sargent, E, Lindquist, R, Schier, G, Hemingsen, C, Wong, J C, Gerhart, J D, Marks, E, Lindqvist |
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| Rok vydání: | 1998 |
| Předmět: |
Multidisciplinary
biology Antibody Affinity Immunoglobulin Variable Region Proteins Biological Sciences medicine.disease_cause Molecular biology Primary and secondary antibodies Affinities Antibodies Mice Antigen biology.protein medicine Animals Humans Genomic library Antigens Antibody Chlamydia trachomatis Gene Single-Chain Antibodies Gene Library |
| Zdroj: | Proceedings of the National Academy of Sciences. 95:6157-6162 |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | A large library of phage-displayed human single-chain Fv antibodies (scFv), containing 6.7 × 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. |
| Databáze: | OpenAIRE |
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