Selection of human anti-human immunodeficiency virus type 1 envelope single-chain antibodies from a peripheral blood cell-based phage repertoire
| Autor: | Marck J. M. Koolen, Piet J. Boender, B. van Gemen, Bert Kazemier, G. van der Groen, Hennie R. Hoogenboom, J. J. W. De Haard, P. Oudshoorn, Jan-Willem Arends |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
medicine.drug_class
viruses Molecular Sequence Data Immunoglobulin Variable Region HIV Infections Biopanning HIV Antibodies HIV Envelope Protein gp120 Biology Monoclonal antibody Binding Competitive Epitope HIV Envelope Protein gp160 Peptide Library Virology medicine Humans Bacteriophages Amino Acid Sequence Immunoglobulin Fragments Linear epitope Panel reactive antibody Molecular biology Primary and secondary antibodies HIV Envelope Protein gp41 HIV-1 Leukocytes Mononuclear biology.protein Hybridoma technology Sequence Analysis Single-Chain Antibodies |
| Zdroj: | Journal of General Virology. 79:2883-2894 |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/0022-1317-79-12-2883 |
| Popis: | Monoclonal antibodies play an important role in the development of diagnostic assays. Instead of using hybridoma technology to isolate human immunodeficiency virus type 1-specific antibodies, a phage-displayed antibody library was generated from a small number (10(7)) of peripheral blood lymphocytes from a seropositive donor. Two families of single-chain antibodies (scFvs) were selected by biopanning with the envelope precursor gp160. ELISA and competition in the BIAcore system revealed that one antibody family recognized a conformation-sensitive epitope within gp120, while the other antibody family was gp41-specific. The latter group had sequence similarity to antibodies recognizing the cluster III epitope of gp41. Binding of scFvs to gp160 could be inhibited with the donor's serum antibodies, indicating that antibodies with a similar specificity were circulating in the donor's blood. Competition experiments suggested that the epitope of the anti-gp41 antibodies was recognized by a broad range of patients' sera: 21 out of 22 sera from North American and all 20 sera from African seropositive patients inhibited binding of scFvs. In contrast, three sera from this panel did not react with the epitope of the anti-gp120 antibodies. These data indicate that, because of the conserved nature of its epitope, the anti-gp41 antibody will be suitable for diagnostic applications. |
| Databáze: | OpenAIRE |
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