Serum N-Glycan Profiling Predicts Prognosis in Patients Undergoing Hemodialysis
Figure 1
General protocol for the integrated glycoblotting technique and workflow for glycoblotting-based high-throughput clinical glycan analysis. Serum samples of 10 μL (a) were applied to the “Sweet Blot” automated machine for glycoblotting. After enzymatic cleaving from serum protein, the total serum N-glycans released in the digest mixture (b) were directly mixed with BlotGlyco H beads (Sumitomo Bakelite, Co., Tokyo, Japan) to capture N-glycans (c). After the beads had been separated from other molecules by washing (d), sialic acid was methyl esterified (e). The processed N-glycans were then labeled with benzyloxyamine (BOA) and released from the BlotGlyco H beads (f). Mass spectra of BOA-labeled N-glycans were acquired with an Ultraflex III instrument (Bruker Daltonics, Germany) (g).
