Fully Automated Sample Preparation Module for Glycan Analysis MUP-3100

Significance of the Topic

Glycan structures on therapeutic antibodies play a critical role in determining stability, efficacy and safety of biopharmaceuticals. Glycoform profiling is an essential quality control parameter in antibody manufacturing, ensuring batch consistency and functional performance in clinical applications.

Objectives and Study Overview

This study introduces the Shimadzu MUP-3100 Fully Automated Sample Preparation Module for Glycan Analysis, developed in collaboration with Sumitomo Bakelite Co., Ltd. combined with the Auto-EZGlyco™ mAb-N Kit and optional O-Glycan Upgrade Kit. The primary aim is to fully automate the complex pretreatment steps—antibody purification, glycan release, fluorescent labeling, and cleanup—to enhance throughput, reproducibility and traceability in glycan profiling workflows.

Methodology and Instrumentation

• The MUP-3100 automates dispensing, reagent addition, heating, centrifugation, solid-phase extraction and sample delivery, using image processing and sensor checks to prevent errors.
• Auto-EZGlyco™ mAb-N Kit supports N-glycan release and labeling; optional O-Glycan Upgrade Kit enables safe O-glycan pretreatment without hazardous reagents.
• Analytical instruments include HPLC with fluorescence detection, LC-MS, Shim-pack GIST-HP Amide [Metal Free] column, LabSolutions™ software and i-PeakFinder™ for automated peak integration.

Key Results and Discussion

• Automated pretreatment processes up to 24 antibody samples in 6 hours versus over two days manually, maintaining stable peak area ratios across plates.
• O-glycan protocol can be completed in 11 hours, replacing toxic hydrazine steps with hydroxylamine chemistry for high recovery and safety.
• Image-based reagent positioning and pipette tip/column grip monitoring ensure consistent workflow and immediate pause on errors, with video capture for troubleshooting.

Benefits and Practical Applications

• Significant labor savings and reduced downtime through unattended operation, including overnight and weekend runs.
• Improved reproducibility by eliminating inter-analyst variability and human errors.
• Enhanced QC data traceability via automated report generation, database storage of pretreatment logs and error videos.
• Enables efficient optimization of cell culture conditions by rapidly profiling antibody glycosylation patterns.

Future Trends and Applications

• Integration with AI-driven analytics and real-time data feedback for adaptive bioprocess control.
• Expansion to multiplexed workflows for other glycoprotein therapeutics and complex glycan classes.
• Miniaturization of sample preparation modules for high-throughput screening and continuous manufacturing environments.
• Potential coupling with online monitoring tools to enable near real-time glycan profiling during bioreactor operation.

Conclusion

The Shimadzu MUP-3100, in tandem with Auto-EZGlyco™ kits, streamlines glycan sample preparation for antibody and glycoprotein analysis, offering rapid, reproducible and traceable workflows that support rigorous quality control and process optimization in biopharmaceutical development.

References

• Kameyama et al. A practical method of liberating O-linked glycans from glycoproteins using hydroxylamine and an organic superbase. Biochemical and Biophysical Research Communications. 513(1):186-192 (2019).