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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Kang, A. S. Barbas, C. F. Gram, H. Lerner, R. A. Marconi, L. A. Collet, T. A. |
| Description | Author Affiliation: Gram H ( Scripps Research Institute, Department of Molecular Biology, La Jolla, CA 92037.); |
| Abstract | We have used a combinatorial immunoglobulin library approach to obtain monoclonal antibodies from nonimmune adult mice, thereby establishing the principles of (i) accessing naive combinatorial antibody libraries for predetermined specificities and (ii) increasing the affinity of the selected antibody binding sites by random mutagenesis. A combinatorial Fab library expressing immunoglobulin mu and kappa light-chain fragments on the surface of filamentous phage was prepared from bone marrow of nonimmunized, adult BALB/c mice with the multivalent display vector pComb8. Phage displaying low affinity Fabs (binding constants, 10(4)-10(5) M-1) binding to a progesterone-bovine serum albumin conjugate were isolated from the library. Random mutagenesis of the heavy- and light-chain variable regions expressed in the mono-valent phage display vector pComb3 was performed by error-prone PCR, and subsequently clones with improved affinity for the hapten conjugate were selected. We demonstrate that antibodies with desirable characteristics from a nonimmune source may be selected and affinity maturation may be achieved by using the twin vectors pComb8 and pComb3, thus opening the route to obtaining specific antibodies from a generic library and bypassing immunization. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 8 |
| Volume Number | 89 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1992-05-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Antibodies, Monoclonal Genetics Bone Marrow Immunology Gene Library Genes, Immunoglobulin Immunoglobulin Fab Fragments Immunoglobulin Variable Region Immunoglobulin Kappa-Chains Immunoglobulin Mu-Chains Amino Acid Sequence Animals Enzyme-Linked Immunosorbent Assay Mice Mice, Inbred BALB C Molecular Sequence Data Mutagenesis Oligodeoxyribonucleotides Plasmids Polymerase Chain Reaction Restriction Mapping Sequence Homology, Nucleic Acid Comparative Study Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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