High-throughput Quantitative Glycomics Enabled by 12-plex Isobaric Multiplex Labeling Reagents for Carbonyl-Containing Compound (SUGAR) Tags

Glycans, which are widely distributed on most proteins and cell surfaces, are a class of important biomolecules playing important roles in various biological processes such as immune response and cellular communication. Modern mass spectrometry (MS) and novel chemical probes together greatly facilitate the routine analysis of glycans. However, the requirement of high-throughput analysis still calls for advanced tools to be developed. Recently, we devised isobaric multiplex reagents for carbonyl-containing compound (SUGAR) tags for N-glycans analysis capable of processing four samples at a time. To further improve the throughput, we utilized the subtle mass differences among different isotopologues combinations and expanded the multiplexing capacity to twelve channels, a three-fold throughput improvement compared to the original setup and achieved ultra-high throughput N-glycans analysis in a single LC-MS/MS injection. We then applied 12-plex SUGAR tags to profile the N-glycans in four subtypes of human Immunoglobulin G (IgG) and to investigate the N-glycans changes in the endometrial cancer cells (ECC1) treated with Atovaquone, a quinone antimicrobial medication, and a dihydroorotate dehydrogenase (DHODH) inhibitor.