Monoclonal Antibody Critical Quality Attribute Verification Using Capillary Scale LC

Monoclonal Antibodies (mAb) have become a major class of medications that are now used around the world. Due to the complex nature of mAbs, a single method for determining structure and function proves challenging. Liquid chromatography (LC) has been used to determine various properties of the antibody required to produce the desired effect(s), known as critical quality attributes (CQAs). Capillary scale LC offers the same ability to determine CQAs as more conventional analytical LC, while considerably reducing the amount of sample and solvent consumed, leading to greener and less expensive separations. Additionally, when coupling to electrospray ionization mass spectrometry (ESI-MS), the lower flow rates of capillary scale separations can enhance ionization efficiency and overall sensitivity of the technique. In this work, the Axcend Focus LC was used to determine various CQAs including intact and fragmented masses, charge variance, glycosylation, and antibody aggregation. These separations showcase the functionality of various capillary scale separation modes including RPLC, IEX, HILIC, and SEC. This marks a considerable step forward in implementing capillary scale separations in the biopharmaceutical industry.