High throughput plasma N-glycome profiling using multiplexed labelling and UPLC with fluorescence detection

Area of Science:

• Glycomics
• Analytical Chemistry
• Biochemistry

Background:

• Glycomic profiling is crucial for understanding biological processes.
• Current methods can be time-consuming, limiting large-scale studies.
• High-throughput analysis is needed for clinical and research applications.

Purpose of the Study:

• To develop a rapid and multiplexed method for N-glycan profiling.
• To enhance sample throughput for plasma glycomic analysis.
• To enable efficient large-scale glycomic studies.

Main Methods:

• N-glycans from plasma samples were individually labeled with specific fluorescent tags (aniline, 2-aminobenzamide, 2-aminoacridone).
• Labeled N-glycans were mixed and co-injected for separation using Hydrophilic Interaction Liquid Chromatography (HILIC) with fluorescence detection.
• The multiplexed method was transferred to Ultra-Performance Liquid Chromatography (UPLC) for increased speed and throughput.

Main Results:

• A single HILIC-fluorescence run allowed simultaneous separation of multiplexed N-glycans.
• Transfer to UPLC-fluorescence increased sample throughput from 24 to 864 plasma samples per day.
• The method provides rapid and efficient glycomic profiling.

Conclusions:

• The developed method offers a significant advancement in the speed and efficiency of plasma N-glycan analysis.
• This high-throughput glycomic profiling technique can facilitate larger clinical and research studies.
• The multiplexed approach enhances analytical capacity for glycomic investigations.