Rapid Intact mass based multi-attribute method in support of mAb upstream process development

Area of Science:

• Biopharmaceutical Development
• Analytical Chemistry
• Protein Chemistry

Background:

• N-linked glycosylation is a critical quality attribute for monoclonal antibodies (mAbs), influencing stability, immunogenicity, and efficacy.
• Bioreactor operational parameters significantly impact mAb glycosylation.
• Timely monitoring of glycosylation during cell culture is crucial for successful process development.

Purpose of the Study:

• To describe a simple, rapid, and robust Multi-Attribute Method (MAM) for analyzing N-linked glycosylation of mAbs.
• To enable direct monitoring of glycosylation profiles from bioreactor samples.
• To confirm the utility of intact mass analysis for critical quality attribute assessment.

Main Methods:

• Development of a Multi-Attribute Method (MAM) based on intact mass analysis.
• Optimization for analyzing light and heavy chains from dithiothreitol (DTT) reduced mAbs.
• Direct sampling from bioreactors for analysis.

Main Results:

• The intact mass MAM effectively monitors N-linked glycosylation profiles, light/heavy chain identity, light chain glycation, and non-glycosylated heavy chain (NGHC).
• N-linked glycosylation profiles correlate well with release glycan assays and peptide-based MAM.
• NGHC results are comparable to capillary gel electrophoresis (CGE) and peptide-based MAM.

Conclusions:

• Intact mass-based MAM provides a rapid and reliable method for monitoring mAb glycosylation during cell culture.
• This approach supports efficient bioprocess development and quality control.
• The method is suitable for open-access instrumentation, enhancing accessibility.