Covalent Labeling/Mass Spectrometry of Monoclonal Antibodies with Diethylpyrocarbonate: Reaction Kinetics for Ensuring Protein Structural Integrity

• DOI: 10.1021/jasms.0c00067
• Publication Date: 2020-04-20T16:24:17Z
• Authors: Patanachai K. Lim...; Bo Zhao; Xiao Pan; Stephen J. Eyles; Richard W. Vachet
• Keywords: Structural integrity

Diethylpyrocarbonate (DEPC)-based covalent labeling together with mass spectrometry is a promising tool for the higher-order structural analysis of antibody therapeutics. Reliable information about structure can be obtained, though, only when protein's integrity preserved during labeling. In this work, we have evaluated applicability DEPC reaction kinetics ensuring monoclonal antibodies (mAbs) By monitoring modification extent selected proteolytic fragments as function concentration, find that common concentration used different in formulated samples without perturbing their structure. Under these conditions, accommodate up to four modifications being structurally perturbed, indicating multidomain proteins withstand more than one label, which contrasts previously studied single-domain proteins. This extensive provides sensitive measure structure, making DEPC-based labeling-mass suitable analyses mAbs.