Forced Degradation Study of Monoclonal Antibody Using Two-Dimensional Liquid Chromatography

摘要: Monoclonal antibody (mAb) products are extraordinarily heterogeneous due to the presence of a variety enzymatic and chemical modifications, such as deamidation, isomerization, oxidation, glycosylation, glycation, terminal cyclization, aggregation, fragmentation. Forced degradation study is common practice assess potential modifications pathways mAbs upon extreme conditions, including light exposure local pH. The degraded samples subject characterization using wide array analyses, ion exchange chromatography (IEC) for charge variants size exclusion (SEC) variants. In this proof-of-concept study, two-dimensional liquid (2D-LC) approach was successfully applied mAb forced evaluation distribution in IEC profile. variant peaks first-dimension (1D) were fractioned by heart-cutting schedule, integrated peak parking feature accommodated later-eluting 1D cuts when previous ones still under second-dimension (2D) analysis. Thus, singly injection, all fractions acquired 2D SEC This demonstrated that compatible with 2D-LC method sensitivity sufficient determine aggregates fragments each individual cut. calculated total stressed mAb-1 comparable those detected one dimensional SEC. Most importantly, IEC-SEC capable bridging real-time efficient manner. It observed enriched most basic region profile, especially highly samples. first application intact analysis per authors’ knowledge.