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

Carolina Lanter1, Malka Lev1, Li Cao1

  • 1CMC Analytical, R&D Medicinal Science & Technology, GSK, 1250 S Collegeville Road, UP1400, Collegeville, PA 19426, USA.

Journal of Biotechnology
|April 16, 2020
PubMed
Summary

A new Multi-Attribute Method (MAM) using intact mass analysis offers rapid N-linked glycosylation profiling for monoclonal antibodies (mAbs) during bioreactor operations. This method ensures critical quality attributes are monitored efficiently for successful process development.

Keywords:
CharacterizationGlycationMass spectrometryMulti-attribute method (MAM)N-linked glycosylationNon-glycosylated heavy chain (NGHC)Upstream process developmentmAb

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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.