Generation of a family-specific phage library of llama single chain antibody fragments that neutralize HIV-1
| Autor: | Willie W. L. Koh, Hans de Haard, Robin A. Weiss, Bart Hoorelbeke, Maria Gonzalez-Pajuelo, Andrea Gorlani, Agnieszka Szynol, Marlén M. I. Aasa-Chapman, Anna Forsman, Theo Verrips, Soren Steffensen |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Phage display
Sequence analysis Immunology Molecular Sequence Data VHH Biology HIV Envelope Protein gp120 Biochemistry Microbiology Neutralization Antibodies Antigens CD4 Retrovirus Phage Display Peptide Library Sequence Homology Nucleic Acid Humans Binding site Peptide library Molecular Biology Llama Heavy Chain Antibodies AIDS Vaccines Binding Sites Base Sequence HIV virus diseases Cell Biology Surface Plasmon Resonance biology.organism_classification Virology Kinetics CD4 Antigens Mutation biology.protein HIV-1 Neutralizing Antibodies Antibody Single-Chain Antibodies Human Immunodeficiency Virus |
| Zdroj: | The Journal of Biological Chemistry |
| Popis: | Recently, we described llama antibody fragments (VHH) that can neutralize human immunodeficiency virus, type 1 (HIV-1). These VHH were obtained after selective elution of phages carrying an immune library raised against gp120 of HIV-1 subtype B/C CN54 with soluble CD4. We describe here a new, family-specific approach to obtain the largest possible diversity of related VHH that compete with soluble CD4 for binding to the HIV-1 envelope glycoprotein. The creation of this family-specific library of homologous VHH has enabled us to isolate phages carrying similar nucleotide sequences as the parental VHH. These VHH displayed varying binding affinities and neutralization phenotypes to a panel of different strains and subtypes of HIV-1. Sequence analysis of the homologs showed that the C-terminal three amino acids of the CDR3 loop were crucial in determining the specificity of these VHH for different subtype C HIV-1 strains. There was a positive correlation between affinity of VHH binding to gp120 of HIV-1 IIIB and the breadth of neutralization of diverse HIV-1 envelopes. The family-specific approach has therefore allowed us to better understand the interaction of the CD4-binding site antibodies with virus strain specificity and has potential use for the bioengineering of antibodies and HIV-1 vaccine development. ispartof: Journal of Biological Chemistry vol:285 issue:25 pages:19116-19124 ispartof: location:United States status: published |
| Databáze: | OpenAIRE |
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