What information can a single quadrupole mass spectrometer provide for biopharmaceutical characterization of monoclonal antibodies?

Importance of the Topic

Glycosylation of monoclonal antibodies impacts stability safety and efficacy in biotherapeutics.
Monitoring glycan distribution is a key quality control metric to ensure consistent manufacturing outcomes.

Objectives and Study Overview

• Evaluate the capability of a single quadrupole mass spectrometer for intact and subunit level glycosylation profiling of trastuzumab.
• Compare relative abundances of glycoforms under reduced quality control tolerances.

Methodology

A 50 microgram sample of trastuzumab was reduced with DTT before quenching with formic acid.
Intact and reduced forms were diluted to 250 ng per microliter in 0.1 percent formic acid.
Data were acquired in positive electrospray mode with scan ranges of m/z 1000 to 3000 for intact and m/z 600 to 2400 for subunits.
Spectra were deconvoluted using vendor software with optimized parameters for mass peak shape.

Used Instrumentation

• Agilent 1290 Infinity II Bio LC system
• Agilent InfinityLab Pro iQ Plus single quadrupole LC/MS
• OpenLab CDS software version 2.8 for data processing

Results and Discussion

The single quadrupole system reliably detected five glycoforms on intact trastuzumab around 148 kilodaltons.
Subunit analysis revealed no glycan on the light chain and four distinct glycoforms on the heavy chain.
Relative abundances from deconvoluted spectra matched expectations and aligned with high resolution platforms.

Benefits and Practical Applications

• Provides a cost effective and robust approach for routine glycosylation monitoring.
• Enables rapid QC decision making with simplified instrumentation.
• Supports process verification without the need for high resolution mass analyzers.

Future Trends and Applications

• Adoption of extended mass range quadrupole systems for broader biomolecule characterization.
• Integration with automated sample preparation and data analysis workflows.
• Expansion to other post translational modifications beyond glycosylation.

Conclusion

The Agilent InfinityLab Pro iQ Plus single quadrupole LC/MS system demonstrates reliable intact and subunit glycosylation profiling of monoclonal antibodies.
It offers a practical alternative for quality control laboratories seeking simplified MS solutions.

References

1. Higel F Seidl A Sorgel F Friess W N glycosylation heterogeneity and its influence on mAb structure function and pharmacokinetics Eur J Pharm Biopharm 2016 100 94 100
2. Wang D Baudys J Bundy J Solano M Keppel T Barr J Comprehensive analysis of glycan component of SARS CoV 2 spike proteins using signature ions triggered EThcD mass spectrometry Anal Chem 2020 92 14730 14739