Evaluating BioAccord LC-MS System Performance for Deployment of a Compact, SmartMS-Enabled Biopharma Solution in Late Stage Development and QC Environment – Intact Mass Analysis

Significance of the Topic

Liquid chromatography–mass spectrometry (LC-MS) is essential for the characterization of biotherapeutics. Intact mass analysis of monoclonal antibodies (mAbs) provides accurate molecular weight data and glycoform distribution, critical for product quality, safety, and regulatory compliance in late-stage development and QC environments.

Study Objectives and Overview

This study assesses the reproducibility of the Waters BioAccord LC-MS system for intact mAb glycoprofiling. Two inter-system experiments (continuous operation over 5.5 days and injections over 90 days) and an intra-system comparison across six instruments were performed.

Methodology

The workflow uses rapid reversed-phase LC separation of a NIST mAb reference standard followed by high-resolution time-of-flight detection. Samples were prepared at 0.2 µg/µL concentration, with 0.8 µg injected onto a BEH C4 column using a 7-minute gradient.

Instrumental Setup

• ACQUITY UPLC I-Class PLUS with TUV or FLR detector
• ACQUITY RDa compact TOF mass detector (400–7000 m/z, 2 Hz scan rate)
• UNIFI Scientific Information System with automated acquisition and MaxEnt deconvolution

Results and Discussion

• Continuous 5.5-day study (1,098 injections): ≤2% RSD for the top five glycoforms.
• Discontinuous 90-day study: ≤8% RSD for glycoform relative abundances.
• Six-system comparison: ≤6% RSD across instruments.
• Mass accuracy consistently within 20–25 ppm.

Benefits and Practical Applications

The BioAccord system provides robust, reproducible data with minimal user intervention, supporting routine implementation of LC-MS assays in regulated and non-regulated labs for mAb quality control.

Future Trends and Opportunities

• Integration with multi-attribute methods for comprehensive protein characterization.
• Advanced automation and AI-driven data analytics to enhance throughput and insight.
• Expansion to antibody–drug conjugates, biosimilars, and other complex modalities.

Conclusion

The BioAccord platform demonstrates high stability and reproducibility for intact mass mAb analysis, meeting industry requirements for late-stage development and QC laboratories.

Reference

1. Rogstad S. et al. J Am Soc Mass Spectrom. 2017;28(5):786–794.
2. Waters application notes and technology briefs (2019–2020).