A universal strategy for development of a method for absolute quantification of therapeutic monoclonal antibodies in biological matrices using differential dimethyl labeling coupled with ultra performance liquid chromatography-tandem mass spectrometry.

Although the strategic use of enzymatic digestion combined with isotope dilution mass spectrometry has been increasingly developed and used for the absolute quantification of therapeutic and endogenous proteins in the biopharmaceutical industry over the past several years, the lack of an appropriate internal standard has become the rate-limiting step in the development of a standardized analytical approach to provide bioanalytical support for both preclinical and clinical studies. In this study, we present a universal strategy for fast development and validation (within 1−2 weeks) of a method for absolute quantification of a therapeutic monoclonal antibody in biological matrices using differential dimethyl labeling coupled with UPLC-MS/MS. Differential dimethyl labeling of tryptic peptides generated from the purified therapeutic monoclonal antibody and those derived from proteins in cynomolgus monkey serum with either d(2)- or d(0)-formaldehyde provided a fast, cost-effective, and standardized approach to...