In-Depth Analysis of N- and O-linked Glycans in a Therapeutic Fc-Fusion Protein Using a Novel MS Data Workflow

CASSS: MS 2022
September 19, 2022

Glycosylation plays a crucial role in protein structure and function, and affects numerous properties of biopharmaceuticals, such as their thermal stability, reactivity with receptors, and immunogenicity. Consequently, thorough characterization of glycosylation is required to ensure a safe and efficacious biopharmaceutical product. Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. Its complexity often necessitates a combination of different sample preparation strategies to achieve complete characterization, resulting in large volumes of complex MS data which typically requires laborious and time-consuming expert analysis.

Here we describe software-driven MS-based methods that provide a consolidated data analysis approach, streamlining the complete characterization of the glycosylation profile of Etanercept.