Glycosylation is the most common post-translational modification (PTM) during the production of mAbs, and its pattern may affect the drug PK, efficacy and safety. For example, high mannose has shown to increase mAb clearance. As such, it is important to monitor glycoform pattern not only in biotherapeutics manufacturing but also follow its fate in vivo over time after administration to humans or animals. An immunocapture-LC/MS assay was developed to assess a variety of glycoforms of a IgG antibody, which includes purification of drug of interest from plasma sample and analysis of glycopeptides by LC-MS/MS after trypsin digestion. Here presented are results from investigation of various glycoforms including G0F, G1F, G2F, Mann5 and sialic acid containing glycans of BI X in human plasma samples. The results showed that the major glycoforms G0F and G1F remained relatively constant over time, Mann5 containing variants showed a faster clearance.
Learning Objectives:
Upon completion, participant will be able to utilize immunocapture-LC/MS technology to assess in vivo critical quality attributes of biologics.
Upon completion, participant will be able to specify the potential effects of critical quality attributes on the biologics
Upon completion, participant will be able to develop different strategies for glycoform analysis