Charge variant profiling is vital for biotherapeutic development, as it tracks molecular attributes that impact drug safety and efficacy. However, analyzing complex biologic modalities such as Fc fusion proteins poses challenges due to their heterogeneity, including extensive glycosylation and sialylation. This case study discusses the strength of online icIEF-UV/MS in identifying problematic acidic variants and post-translational modifications, enabling risk mitigation and a deeper mechanistic understanding of highly glycosylated therapeutics.

Key takeaways from this presentation:

• Online icIEF-UV/MS enables rapid and robust analysis of charge variants, even in highly heterogeneous proteins
• Enzymatic treatments and optimized conditions improve resolution and help identify post-translational modifications and glycan-related variants
• This approach is a valuable tool for accelerating biologics pipeline development and in-depth characterization
• Rapid identification of PTMs enables risk mitigation in developing highly glycosylated therapeutics