Given the complexity of producing bispecific antibodies, suitable analytical methods to detect and measure the levels of undesired variants are essential. Here, a novel MS-based method for variant detection is presented.
Long-term column use can lead to on-column methionine oxidation during LC–MS/MS peptide mapping of antibody-based biotherapeutics. Following the approach described here minimizes the risk of measuring oxidative artifacts, and helps generate high quality data to provide reliable quantitative information about product-related heterogeneities.
Applications of size-exclusion chromatography (SEC) are presented for characterization and quality control of novel biotherapeutic products,
including antibody–drug conjugates, hydrophobic proteins, and coformulations.
In this study, a conjugation strategy for Fab–ferritin conjugates, used for drug delivery, was successfully optimized using LC–MS. Characterization of the resulting conjugates was performed using SEC-MALS-QELS.
The potential of multidimensional online peptide mapping analysis as a strategy for improving a postlabeling workflow for protein–protein interactions is demonstrated using both hydroxy radical footprinting–mass spectrometry (HRF–MS) and LC–MS/MS.
A simple and robust size-exclusion chromatography (SEC) method has been developed for characterizing a multimeric PEG–protein conjugate. A wide range of size variants of the conjugate, ranging from 50 kDa to >1000 kDa, can be resolved and quantitated.