As a continuation of our work on boronic acid lectin affinity chromatography (BLAC), in this paper we introduce an automated
affinity micropartitioning approach using combined boronic acid and concanavalin A (BLAC/Con A) resin-filled micropipette
tips to isolate and enrich human serum glycoproteins. The
N-linked oligosaccharides of the partitioned glycoproteins were removed by PNGase F enzyme digestion, followed by 8-aminopyrene-1,3,6-trisulfonic
acid labeling. Capillary gel electrophoresis with blue LED-induced fluorescence detection was applied in a multiplexed format
for comparative glycan profiling. The efficiency of BLAC affinity micropartitioning was compared with that of the individual
lectin and pseudolectin affinity enrichment. Finally, we report on our findings in glycosylation differences in human serum
samples from healthy and prostate cancer patients by applying BLAC/Con A micropipette tip-based enrichment and comparative
multicapillary gel electrophoresis analysis of the released and labeled glycans.
Figure Fluorophore labeling and purification in BLAC–mCE based glycosylation profiling
Keywords Affinity chromatography - Capillary gel electrophoresis - Glycan profiling
Alex Monzo and Marcell Olajos contributed equally to this work.