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Rapid and comprehensive monoclonal antibody Characterization using microfluidic CE-MS.

Li Cao1, Daniel Fabry1, Kevin Lan1

  • 1CMC Analytical, GlaxoSmithKline, 1250 S. Collegeville Road, UP 1400, Collegeville, PA, 19426, USA.

Journal of Pharmaceutical and Biomedical Analysis
|July 15, 2021
PubMed
Summary

Rapid peptide mapping and charge variant analysis using microfluidic CE-MS offer a high-throughput method for monitoring monoclonal antibody (mAb) critical quality attributes (CQAs) in biopharmaceutical development.

Keywords:
charge variantsmicrofluidic CE-MSmonoclonal antibody (mAb)native intact analysispeptide mappingpost-translation modifications (PTMs)

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Area of Science:

  • Biopharmaceutical analysis
  • Analytical chemistry
  • Quality by Design (QbD)

Background:

  • Monoclonal antibody (mAb) critical quality attributes (CQAs) are essential for drug safety and efficacy.
  • Current methods for CQA analysis can be time-consuming and resource-intensive.
  • Quality by Design (QbD) principles necessitate robust and efficient analytical strategies.

Purpose of the Study:

  • To develop and validate rapid peptide mapping and native intact charge variant analysis methods.
  • To comprehensively characterize and monitor mAb CQAs using a microfluidic capillary electrophoresis-mass spectrometry (CE-MS) platform.
  • To demonstrate the potential of these methods for supporting biopharmaceutical development.

Main Methods:

  • Development of ultrafast peptide mapping using microfluidic CE-MS.
  • Implementation of native intact charge variant analysis via microfluidic CE-MS.
  • Characterization of CQAs including primary structure, post-translational modifications, and glycosylation.
  • Assignment of charge variant identities using mass shifts, peptide mapping data, and electrophoretic mobility.
  • Model simulation to correlate peptide-level and intact-level data.

Main Results:

  • Peptide mapping provided comprehensive CQA analysis comparable to conventional LC-MS.
  • Native intact analysis resolved mAb charge variants with high resolution, similar to cIEF.
  • Charge variant identities and major glycoforms were successfully assigned.
  • Model simulation confirmed high correlation between peptide mapping and intact mass spectrometry data.
  • The developed methods demonstrated high throughput and consistency.

Conclusions:

  • Microfluidic CE-MS enables rapid and comprehensive characterization of mAb CQAs.
  • These methods provide a powerful tool for supporting biopharmaceutical development and quality control.
  • The integrated approach enhances understanding and control of mAb product quality.