SmartMS-ENABLED LC-MS SYSTEM FOR BIOTHERAPEUTIC DEVELOPMENT IN REGULATED/NON-REGULATED ENVIRONMENTS

Significance of the Topic

Liquid chromatography–mass spectrometry (LC-MS) plays a pivotal role in characterizing biotherapeutics by providing detailed molecular information on intact proteins, peptides and glycans. However, its adoption in routine laboratories is often limited by complex workflows and the need for specialized expertise. The SmartMS-enabled BioAccord system addresses these challenges with a user-friendly interface, automated methods and compact footprint, enabling consistent performance in both regulated and non-regulated environments.

Objectives and Study Overview

This work demonstrates the performance of the BioAccord LC-MS platform using case studies on antibody-drug conjugates (ADCs) and monoclonal antibodies (mAbs). Key aims include:

• Intact protein analysis to determine drug-to-antibody ratios (DAR) by SEC-MS
• Peptide mapping for forced degradation assessment
• Evaluation of inter- and intra-system reproducibility over three months

Methodology and Instrumentation

• Instrument setup: ACQUITY UPLC I-Class with ACQUITY RDa Detector, coupled to UNIFI Scientific Information System 1.9.4
• Intact protein analysis: Native SEC-UV-MS and denaturing SEC-UV-MS using Protein BEH SEC/C4 columns; full scan ESI+ mode (m/z 400-7000)
• Peptide mapping: Tryptic digest of mAb samples after forced light degradation; separation on XSelect CSH C18 column; MS/MS fragmentation (m/z 50-2000)
• Performance evaluation: Humanized mAb mass check standard run across six BioAccord systems over 90 days; assessment of glycoform %RSD for reproducibility

Main Results and Discussion

• Intact Protein Analysis: DAR values for cysteine-conjugated ADCs were consistent with in-house methods (HIC and Q-TOF systems), confirming accuracy of automated workflows.
• Peptide Analysis: Forced degradation of mAb samples revealed increased levels of specific modifications; extracted ion chromatograms and high-energy fragmentation validated peptide identifications and quantitation.
• Instrument Performance: Glycoform abundance measurements (G0F/G0F to G2F/G2F) showed <10% %RSD both intra- and inter-system, demonstrating robust reproducibility.

Benefits and Practical Applications

The BioAccord platform offers:

• An intuitive, workflow-driven interface reducing training time
• Compliance-ready software supporting regulated environments
• Automated acquisition, processing and reporting across intact, subunit, peptide and glycan analyses
• Compact footprint suitable for diverse laboratory settings

Future Trends and Opportunities

• Expansion of SmartMS workflows to include real-time data analytics and AI-assisted interpretation
• Integration with laboratory information management systems for seamless data tracking
• Development of higher-throughput methods to support large-scale biotherapeutic screening
• Integration of multi-omics capabilities for deeper characterization

Conclusion

The SmartMS-enabled BioAccord system simplifies LC-MS analyses for biotherapeutics, delivering performance on par with established methods while improving ease of use and reproducibility. Its automated workflows and compliance features make it a valuable tool for routine QC, R&D and regulated laboratory environments.