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pmid:37526474
Determination of Protein Monoclonal-Antibody Epitopes by a Combination of Structural Proteomics Methods.
Evgeniy V Petrotchenko · Elisabete M Nascimento · Jody Melton Witt · Christoph H Borchers
Journal of proteome research · 2023 doi:10.1021/acs.jproteome.3c00159
Paper
complete
Experiment
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Data
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Analysis
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Execution
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Complete file
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Each technique provides orthogonal structural information on the structure and the location of protein interaction sites. Here, we have characterized a monoclonal antibody epitope for a protein antigen by a combination of differential photoreactive surface modification (SM), cross-linking (CL), differential hydrogen-deuterium exchange (HDX), and epitope extraction/excision. We found that experimental data from different approaches agree with each other in determining the epitope of the monoclonal antibody on the protein antigens using the HIV-1 p24-mAb E complex as an illustrative example. A combination of these multiple structural proteomics approaches results in a detailed picture of the interaction of the proteins and increases confidence in the determination of the final structure of the protein interaction interface. 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