title: Rapid quantitative profiling of N-glycan by the glycan-labeling method using 3-aminoquinoline/α-cyano-4-hydroxycinnamic acid.
abstract: Protein glycosylation is a crucial phenomenon for understanding protein functions, since its patterns and degree are associated with many biological processes, such as intercellular signaling and immune response. We previously reported a novel glycan-labeling method using a 3-ainoquinoline/α-cyano-4-hydroxycinnamic acid (3-AQ/CHCA) liquid matrix for highly sensitive detection by matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry (MS). In the present study, we examined the practicality of this method for qualitative and quantitative glycan profile analysis. We first investigated the reproducibility of the data for 16 N-glycans prepared from human epidermal growth factor receptor type 2 (HER2). All of the data obtained in intra-assays and interassays were highly correlated with statistical significance (R(2) > 0.9, p < 0.05). In addition, the HER2 glycosylation pattern differed significantly between different breast cancer cell lines SK-BR-3 and BT474 in a comparative analysis of profile data. Finally, the quantitative capability of this method was examined by using PA-labeled monosialylated N-glycan as an internal standard (IS). Using IS for AQ-labeled neutral and sialylated standard glycans, the ion peak intensity was highly linear (R(2) > 0.9) from 0.5 to 5000 fmol. Furthermore, using IS for HER2 N-glycans, all of the N-glycans were highly linear with their dilution factors (R(2) > 0.9). These results suggest that our developed AQ labeling method enabled rapid qualitative and quantitative analyses of glycans. This glycan analysis method should contribute to the field of biomarker discovery and biomedicine in applications such as quality control of biotechnology-based drugs.
authors: Koichi Tanaka Laboratory of Advanced Science and Technology, Shimadzu Corporation, 1, Nishinokyo-Kuwabaracho Nakagyo-ku, Kyoto 604-8511, Japan. kaoru07@shimadzu.co.jp
authors: Kaoru
authors: K
authors: Kaneshiro
authors: Makoto
authors: M
authors: Watanabe
authors: Kazuya
authors: K
authors: Terasawa
authors: Hiromasa
authors: H
authors: Uchimura
authors: Yuko
authors: Y
authors: Fukuyama
authors: Shinichi
authors: S
authors: Iwamoto
authors: Taka-Aki
authors: TA
authors: Sato
authors: Kazuharu
authors: K
authors: Shimizu
authors: Gozoh
authors: G
authors: Tsujimoto
authors: Koichi
authors: K
authors: Tanaka
mesh_terms: D000634:Aminoquinolines / Q000737:chemistry*; D045744:Cell Line, Tumor; D003373:Coumaric Acids / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D000634:Aminoquinolines; D003373:Coumaric Acids; D011134:Polysaccharides; D018719:Erb-b2 Receptor Tyrosine Kinases; C007175:alpha-cyano-4-hydroxycinnamate