A rapid and high-throughput biotechnology quantifies major glycoforms of human IgG1 mAb drugs.

Human IgG1 monoclonal antibody (mAb) drugs are widely manufactured using mammalian cells and contain three major glycoforms on asparagine residue at position 297 (Asn297) in their Fc region: fucosylated, afucosylated, and high-mannose glycans. The N-glycans at Asn297 directly impact IgG1 mAb efficacy through effector functions and half-life. Thus, N-glycosylation is a critical quality attribute for IgG1 mAb therapeutics and needs to be controlled during manufacturing and release to ensure lot-to-lot consistency and demonstrate similarity for IgG1 mAbs and biosimilars. In this study, we generate two mouse mAbs, mAb-NG and mAb-G, that specifically recognize nonglycosylated and glycosylated human IgG1, respectively. Using mAb-G, we establish a technology in a biolayer interferometry format with high-throughput potential that rapidly quantifies the relative amount (%) of each glycoform of IgG1 mAbs, either purified or in harvest media. This biotechnology can accelerate cell line and biomanufacturing process development and potentially serve as a component of process analytical technology for IgG1 mAb drugs.