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Research

BlotGlycoABC^TM, an Integrated Glycoblotting Technique for Rapid and Large Scale Clinical Glycomics ^*,S

Yoshiaki Miura, Megumi Hato, Yasuro Shinohara, Hiromitsu Kuramoto, Jun-ichi Furukawa, Masaki Kurogochi, Hideyuki Shimaoka, Mitsuhiro Tada^¶, Kazuaki Nakanishi^||, Michitaka Ozaki^||, Satoru Todo^|| and Shin-Ichiro Nishimura^,**

From the Graduate School of Life Science and Frontier Research Center for Post-Genomic Science and Technology, Hokkaido University, Sapporo 001-0021, Japan, Sumitomo Bakelite Co., Kobe 651-2241, Japan, ^¶ Division of Cancer-Related Genes, Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan, and ^|| Department of General Surgery, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan

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^TM 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.