Qualification of a Quantitative Method for Monitoring Aspartate Isomerization of a Monoclonal Antibody by Focused Peptide Mapping

摘要: Aspartate (Asp) isomerization is a common post-translational modification of recombinant therapeutic proteins that can occur during manufacturing, storage, or administration. Asp in the complementarity-determining regions monoclonal antibody may affect target binding and thus sufficiently robust quality control method for routine monitoring desirable. In this work, we utilized liquid chromatography-mass spectrometry (LC/MS)-based approach to identify antibody. To quantitate site-specific antibody, UV detection-based quantitation assay utilizing same LC platform was developed. The qualified implemented product-specific modification. Compared with existing methods, analytical paradigm applicable (or other modifications) subsequently develop rapid, ensure product quality. This first identifies locates product-related impurity (a critical attribute) caused by isomerization, deamidation, oxidation, modifications, then utilizes synthetic peptides MS assist development LC-UV-based chromatographic separates quantifies impurities peaks. established LC-UV has acceptable peak specificity, precision, linearity, accuracy; it be validated used good manufacturing practice environment lot release stability testing. Lay abstract manufacture process storage. Isomerization (CDRs) A (mAb-A) been detected shown have impact on affinity antigen. mass spectrometry-based peptide mapping detect CDRs mAb-A. routinely monitor CDR mAb-A, focused reversed phase separation detection developed qualified. generally modifications specific high-throughput mode