Rapid Conformational Epitope Mapping of Anti-gp120 Antibodies with a Designed Mutant Panel Displayed on Yeast
| Autor: | Hanna Zhu, Barry Kriegsman, Jordi Mata-Fink, Hui Xin Yu, Darrell J. Irvine, K. Dane Wittrup, Melissa C. Hanson |
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| Přispěvatelé: | Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Chemical Engineering, Massachusetts Institute of Technology. Department of Materials Science and Engineering, Ragon Institute of MGH, MIT and Harvard, Koch Institute for Integrative Cancer Research at MIT, Mata-Fink, Jordi, Kriegsman, Barry, Yu, Hui Xin, Zhu, Hanna, Hanson, Melissa C., Irvine, Darrell J., Wittrup, Karl Dane |
| Rok vydání: | 2013 |
| Předmět: |
Immunogen
Protein Conformation Saccharomyces cerevisiae HIV Antibodies HIV Envelope Protein gp120 Biology Yeast display Kidney Protein Engineering Article Epitope Epitopes Peptide Library Structural Biology Humans Binding site Peptide library Molecular Biology Cells Cultured Linear epitope Flow Cytometry Antibodies Neutralizing Virology Molecular biology Antibodies Anti-Idiotypic Epitope mapping Mutagenesis Drug Design CD4 Antigens Mutant Proteins Epitope Mapping Conformational epitope |
| Zdroj: | PMC |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2012.11.010 |
| Popis: | gp120 is a substrate for protein engineering both for human immunodeficiency virus (HIV) immunogen design and as a bait for isolating anti-HIV antibodies from patient samples. In this work, we describe the display of a stripped core gp120 on the yeast cell surface. Validation against a panel of neutralizing antibodies confirms that yeast-displayed gp120 presents the CD4 binding site in the correct conformation. We map the epitope of the broadly neutralizing anti-gp120 antibody VRC01 using both a random mutagenesis library and a defined mutant panel and find that the resultant epitope maps are consistent with one another and with the crystallographically identified contact residues. Mapping the VRC01-competitive antibodies b12 and b13 reveals energetic differences in their epitopes that are not obvious from existing crystal structures. These data suggest mutation sets that abrogate binding to broadly neutralizing antibodies with greater specificity than the canonical mutation D368R, useful in rapidly assessing the nature of a vaccine response. Ragon Institute of MGH, MIT and Harvard |
| Databáze: | OpenAIRE |
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