BioAccord™ LC-MS System Enhancements for Improved Native MS Analysis

Importance of the Topic

BioAccord LC-MS System extensions are crucial for analyzing large protein complexes and conjugates in biopharmaceutical development. Extending the mass detection range up to m/z 9000 enhances native mass spectrometry capabilities, addressing industry needs for advanced protein characterization, quality control, and monitoring of novel modalities.

Study Objectives and Overview

This application brief evaluates recent upgrades to the BioAccord system, focusing on the extended mass range of the ACQUITY RDa Detector. The study compares standard high mass range mode (m/z 400–7000) with the new extended mass range (m/z 400–9000) for native SEC-MS analysis of multimeric proteins and stressed monoclonal antibody samples.

Methodology and Instrumentation

• Chromatography: ACQUITY UPLC Protein BEH SEC Column, 200 Å, 1.7 μm, 2.1 × 150 mm, isocratic 50 mM ammonium acetate at pH 6.8, 30 °C, 10 min.
• Mass Spectrometry: BioAccord LC-MS System with ACQUITY RDa Detector in positive ESI native mode; comparison of HMR (m/z 400–7000) and EMR (m/z 400–9000).
• Calibration: Standard BioAccord calibration solution.

Key Results and Discussion

Analysis of yeast alcohol dehydrogenase revealed dimer and tetramer species in HMR mode, while the EMR mode additionally detected the octamer species (~296 kDa), demonstrating enhanced detection of larger noncovalent complexes. In a freeze/thaw stressed infliximab sample, the extended range enabled observation of a high molecular weight dimer in the m/z 6500–8500 region, which was not detectable in HMR mode. These results confirm the EMR mode’s capability to capture larger protein assemblies and conjugates under native conditions.

Benefits and Practical Applications

• Expanded mass range allows detection of higher-order protein structures and aggregates.
• Improved characterization of antibody drug conjugates and complex biotherapeutic modalities.
• Enhanced quality control and attribute monitoring in biopharmaceutical workflows.

Future Trends and Opportunities

Further developments may include integration with additional nondenaturing separation techniques, deeper analysis of large viral vectors and oligonucleotide conjugates, and automation improvements for high-throughput native MS workflows. Expanding detector sensitivity and mass range could support emerging modalities such as nanoparticle-based therapeutics.

Conclusion

The extension of the BioAccord LC-MS System mass range to m/z 9000 significantly improves native MS analysis of large protein complexes and conjugates. This enhancement addresses evolving demands in biopharmaceutical research and quality control, facilitating robust characterization of increasingly complex modalities.

References

1. Mallis CS, Zheng X, Qiu X, et al. Development of Native MS Capabilities on an Extended Mass Range Q-TOF MS. Int J Mass Spectrom. 2020;458:116451.
2. Wang Y, Schubert M, Ingendoh A, Franzen J. Analysis of non-covalent protein complexes up to 290 kDa using electrospray ionization and ion trap mass spectrometry. Rapid Commun Mass Spectrom. 2000;14(1):12–17.