12-Plex Isobaric Multiplex Labeling Reagents for Carbonyl-Containing Compound (SUGAR) Tag-Enabled High-Throughput...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Molecular Biology

Background:

• Glycans are vital biomolecules involved in cellular recognition and communication.
• Mass spectrometry (MS) and chemical labeling have advanced glycan analysis.
• High-throughput, quantitative glycan analysis requires improved methodologies.

Purpose of the Study:

• To enhance throughput for N-glycan analysis.
• To develop a multiplexed quantitative glycomics strategy.
• To present detailed methods for 12-plex SUGAR tag synthesis and application.

Main Methods:

• Synthesis of 12-plex SUGAR (carbonyl-containing compound) isobaric tags.
• Release and labeling of N-glycans from proteins.
• High-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.
• Data processing for multiplexed quantitative glycomics.

Main Results:

• Expansion of SUGAR tags from 4-plex to 12-plex using a mass-defect strategy.
• Achieved a threefold increase in throughput compared to previous methods.
• Enabled high-throughput N-glycan analysis in a single LC-MS/MS injection.

Conclusions:

• The 12-plex SUGAR tag strategy significantly improves throughput for N-glycan quantification.
• This method facilitates robust and high-throughput quantitative glycomics.
• Detailed protocols are provided for synthesis, labeling, analysis, and data processing.