Many protein-based biopharmaceuticals are glycosylated proteins. Consistent glycosylation is a sensitive marker for a well-controlled process. For this reason, uncontrolled changes in protein glycosylation are of great regulatory concern. The relative amounts of the individual glycan structures need to be monitored during process development to establish the stability of the growth and purification steps of manufacturing. The same measurements are required in the development of formulations and stability testing.

Recent advances in chromatographic approaches utilizing UPLC^® lead to better resolution, sensitivity, and speed for greater reliability in the qualitative and quantitative analysis of protein glycosylation.

The Waters UPLC Glycan Analysis Solution meets the needs of these applications. The ACQUITY UPLC^® BEH Glycan Separation Technology column is optimized for use with the ACQUITY UPLC System with fluorescence detection (FLR), separating the released glycans of biopharmaceuticals as their 2-aminobenzamide (2-AB) derivatives.

UPLC Glycan Analysis Solution:

• ACQUITY UPLC System
• ACQUITY UPLC Fluorescence (FLR) Detector
• UPLC BEH Glycan Technology Column, specifically developed and QC tested to provide superior UPLC component resolution for a range of glycan structures
• Instrument control software with MassLynx^TM or Empower^TM software

The design and development of the UPLC Glycan Separation Technology Column was targeted to the analysis of the glycans that are commonly found or expected on biopharmaceutical proteins, including high mannose, complex, hybrid, and sialylated glycans.

The ACQUITY UPLC FLR Detector is a high-sensitivity, multi-channel fluorescence detector optimized for use with the ACQUITY UPLC System. Innovative flow cell design, low noise electronics, and support for high-speed data rates result in a detector that brings the ability to effectively acquire and quantitate the high-efficiency separations afforded by UPLC.

When combined for use in a biopharmaceutical or pharmaceutical laboratory to monitor glycan profiles, the UPLC Glycan Separation Solution provides robust, high-resolving, reproducible, and rapid methods that out-perform analyses by HPLC.