title: BlotGlycoABCTM, an integrated glycoblotting technique for rapid and large scale clinical glycomics.
abstract: Recent progress in mass spectrometry has led to new challenges in glycomics, including the development of rapid glycan enrichment techniques. A facile technique for exploration of a carbohydrate-related biomarker is important because proteomics research targets glycosylation, a posttranslational modification. Here we report an "all-in-one" protocol for high throughput clinical glycomics. This new technique integrates glycoblotting-based glycan enrichment onto the BlotGlycoABC bead, on-bead stabilization of sialic acids, and fluorescent labeling of oligosaccharides in a single workflow on a multiwell filter plate. The advantage of this protocol and MALDI-TOF MS was demonstrated through differentiation of serum N-glycan profiles of subjects with congenital disorders of glycosylation and hepatocellular carcinoma and healthy donors. The method also permitted total cellular glycomics analysis of human prostate cancer cells and normal human prostate epithelial cells. These results demonstrate the potentials of glycan enrichment/processing for biomarker discovery.
authors.affiliation: Graduate School of Life Science and Frontier Research Center for Post-Genomic Science and Technology, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Miura
authors.forename: Megumi
authors.initials: M
authors.lastname: Hato
authors.forename: Yasuro
authors.initials: Y
authors.lastname: Shinohara
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Kuramoto
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Furukawa
authors.forename: Masaki
authors.initials: M
authors.lastname: Kurogochi
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Shimaoka
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Tada
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Nakanishi
authors.forename: Michitaka
authors.initials: M
authors.lastname: Ozaki
authors.forename: Satoru
authors.initials: S
authors.lastname: Todo
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000737:chemistry / Q000175:diagnosis; D003937:Diagnosis, Differential; D054794:Glycomics / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood / Q000737:chemistry / Q000175:diagnosis; D008297:Male; D008659:Metabolic Diseases / Q000378:metabolism; D011134:Polysaccharides / Q000097:blood / Q000737:chemistry / Q000145:classification; D011471:Prostatic Neoplasms / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D011134:Polysaccharides