Epitope Mapping with Diethylpyrocarbonate Covalent Labeling-Mass Spectrometry.

ABSTRACT: Antigen-antibody epitope mapping is essential for understanding binding mechanisms and developing new protein therapeutics. In this study, we investigate diethylpyrocarbonate (DEPC) covalent labeling-mass spectrometry as a means of analyzing antigen-antibody interactions using the well-characterized model system of TNFα in complex with three different antibodies. Results show that residues buried in the epitope undergo substantial decreases in labeling, as expected. Interestingly, serine, threonine, and tyrosine residues at the edges of the epitope undergo unexpected increases in labeling. The increased labeling of these weakly nucleophilic residues is caused by the formation of hydrophobic pockets upon antibody binding that presumably increase local DEPC concentrations. Residues that are

SUBMITTER: Tremblay CY 

PROVIDER: S-EPMC8785103 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature