Extending the Analytics of Biopharmaceutical QA/QC Labs with the ACQUITY™ QDa™ II Mass Detector

Importance of the Topic

Mass spectrometry increasingly underpins quality control in biopharmaceutical manufacturing. Traditional UV detection often lacks the sensitivity and specificity required to monitor low-level impurities and critical quality attributes in monoclonal antibody products. Compact, robust mass detectors that integrate seamlessly into regulated workflows are essential for ensuring product safety and compliance.

Study Objectives and Overview

This study evaluates the performance and utility of the ACQUITY QDa II Mass Detector in biopharmaceutical QA/QC laboratories. The primary aims were to:

• Assess assay sensitivity, specificity, and reproducibility compared to UV methods
• Demonstrate identity testing of mAb digestion products via complementary determining region (CDR) peptides
• Detect low-abundance impurities such as oxidation and deamidation variants
• Examine integration with Empower 3 Software and benefits for streamlined QC operations

Methodology and Used Instrumentation

Samples were prepared from a Waters mAb Tryptic Digestion Standard at 0.4 mg/mL. Liquid chromatography was performed on an ACQUITY Premier System with a Premier Peptide CSH C18 column (2.1×100 mm, 1.7 μm) at 60 °C, using a gradient of water and acetonitrile (0.1% formic acid) at 0.2 mL/min. The ACQUITY QDa II Mass Detector operated in positive electrospray mode (350–1500 m/z, 5 Hz) with a probe temperature of 600 °C, capillary voltage 1.5 kV, cone voltage 20 V. Empower 3.8.1 controlled full-scan and selected ion recording (SIR) acquisitions.

Main Results and Discussion

The QDa II detector reliably identified CDR peptides for mAb identity testing and delivered up to a 50% higher signal for ions above 1250 m/z compared to its predecessor. In impurity screening, oxidation of the T21 peptide was quantified down to 0.01% (v/v) with a signal-to-noise ratio of ~3, outperforming conventional UV limits. A stability study monitored T21 oxidation and T37 deamidation over 84 hours: oxidation remained stable in sealed vials but increased significantly after septum puncture, while deamidation levels showed no significant change. Relative standard deviations for key CQAs were ≤5%, demonstrating reproducibility and suitability for automated workflows.

Benefits and Practical Applications

• Enhanced sensitivity for larger peptides enables robust CDR mapping and identity confirmation
• Low-level impurity detection meets and exceeds regulatory thresholds (0.01% vs. 0.05%)
• Integrated Empower 3 control streamlines QC method development and reduces operating costs
• Compact, turnkey design supports rapid deployment in cGMP-regulated labs

Future Trends and Applications

Future developments will focus on deeper automation, AI-driven data interpretation, and integration with laboratory information management systems. Expanded multi-attribute monitoring, high-throughput screening platforms, and digital twin models for process monitoring will leverage compact MS detectors. Continued miniaturization and sustainable design will further reduce environmental impact and operational expenses.

Conclusion

The ACQUITY QDa II Mass Detector provides biopharmaceutical QA/QC laboratories with a sensitive, specific, and user-friendly mass spectrometry solution. Its extended mass range, low-level impurity detection, and seamless Empower 3 integration enable accurate identity testing and CQA monitoring under cGMP, enhancing compliance and operational efficiency.

References

1. Ecker DM, Jones SD, Levine HL. The Therapeutic Monoclonal Antibody Market. mAbs. 2015;7(1):9–14.
2. Title 21 C.F.R. Part 210—Current Good Manufacturing Practice in Manufacturing, Processing, Packing, or Holding of Drugs; General.
3. Dong Q, Liang Y, Yan X, et al. The NISTmAb Tryptic Peptide Spectral Library for Monoclonal Antibody Characterization. mAbs. 2018;10(3):354–369.