Automating Rapid High-throughput mAb Attribute Screening of Microbioreactor Cell Culture Media Samples

Importance of the Topic

Monoclonal antibodies (mAbs) are critical biotherapeutics whose development depends on rapid and reliable analytical screening of cell culture media. High-throughput platforms accelerate clone selection, reduce development timelines and enhance data consistency.

Study Objectives and Overview

This work presents an end-to-end automated workflow for mAb attribute screening directly from microbioreactor cell culture samples. Key goals included reducing sample volume requirements, improving throughput and integrating purification, subunit preparation and LC-MS analysis into a single automated platform.

Methodology and Instrumentation

Sample preparation combined Protein A magnetic bead purification, reduction and optional enzymatic digestion for subunit or peptide/glycan analysis. A compact Andrew+ pipetting robot orchestrated 48 samples in a 2.5-hour run using Domino modules for tip handling, plate heating/shaking and magnetic separation.

• Purification reagents: Promega Magne™ Protein A beads, glycine-HCl elution and neutralization buffers.
• Reduction: DTT in 6 M Guanidine-HCl and Tris-HCl.
• Enzymatic digestion: IdeS enzyme for subunit generation.
• LC-MS platform: ACQUITY UPLC I-Class Plus with BioAccord™ TOF MS, BioResolve™ RP mAb Polyphenyl column (2.1×100 mm, 2.7 µm, 450 Å).
• Chromatography: 4.5 min gradient, 0.4 mL/min, 80 °C column, 10 °C sample tray, 3 µL injection.
• Mass spectrometry: ESI+ mode, 50–2000 m/z, 2 Hz scan rate, waters-connect™ lockmass.

Main Results and Discussion

Automated purification delivered reproducible subunit recovery (mass deviations <0.06 Da). LC-MS analysis required 5 min per sample, achieving consistent Fd’ and Fc/2 masses within ±0.3 Da. Glycan profiling showed relative abundance reproducibility across common glycoforms. Overall, the platform processed 48 unpurified or Protein A-purified samples in under 3 hours with minimal user intervention.

Benefits and Practical Applications

The automated workflow offers:

• Rapid turnaround: 2.5 h for sample prep and 5 min per analysis.
• Low sample volume (20–100 µL) compatible with microbioreactors.
• High reproducibility and data reliability via standardized robotics.
• Scalability to subunit, peptide or glycan attribute screening during lead selection.

Future Trends and Opportunities for Application

Future developments may integrate multi-attribute method capabilities, real-time process monitoring via PAT, and advanced software analytics for automated decision-making. Expansion to intact mass profiling, peptide mapping and glycan structure elucidation could further streamline bioprocess development.

Conclusion

This study demonstrates a fully automated, high-throughput mAb attribute screening platform that combines minimal sample volumes, rapid LC-MS analysis and robust reproducibility. Such integration supports accelerated biotherapeutic development and consistent quality control.

References

1. Aghayee A.; Htet Y.; Morrison L.; Koza S.M.; Yu Y.Q. Waters Corporation April 2021.
2. Htet Y.; Koza S.M.; Chen W. Waters Corporation August 2021.
3. Dong J.; et al. Analytical Chemistry. 2016, 88, 5673–5687.