Background:Atypical peaks were observed in capillary electrophoresis with sodium
dodecyl sulfate (CE-SDS) during the development of therapeutic monoclonal IgG4 antibodies
(mAb-X). Based on the previous literature reports, the atypical peak may be caused by various factors
such as post-translational modifications (PTMs), method-induced artifacts, sample degradation
and sequence variants. Due to the high complexity structure of mAbs and the limitations of
CE-SDS, acquiring comprehensive profiling of atypical peaks has historically been challenging.Objective:Here we developed a strategy utilizing complementary analytical methods to identify
the cause of atypical peak.Methods:This strategy includes optimizing reduced CE-SDS method to evaluate artifacts induced
by the analytical method, excluding potential glycation modifications, and utilizing Liquid Chromatograph
Mass Spectrometer (LC-MS) to characterize mAb-X.Results:Our study demonstrates that the atypical peaks of mAb-X are a mixture of method-induced
artifacts and variants in the C-terminal extension sequence of the light chain.Conclusion:Strategy for complementary analytical methods tools helps to identify the cause of
unknown species and plays a key role in product and process characterization.