Hyphenated Technique for Releasing and MALDI MS Analysis of O-Glycans in Mucin-Type Glycoprotein Samples

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• 作者：Keita Yamada ; Satomi Hyodo ; Mitsuhiro Kinoshita ; Takao Hayakawa ; Kazuaki Kakehi
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：September 1, 2010
• 年：2010
• 卷：82
• 期：17
• 页码：7436-7443
• 全文大小：343K
• 年卷期：v.82,no.17(September 1, 2010)
• ISSN：1520-6882

文摘

We developed an automatic apparatus for the release of O-glycans from mucin-type glycoproteins and proteoglycans (Matsuno, Y.-k.; Yamada, K.; Tanabe, A.; Kinoshita, M.; Maruyama, S.-z.; Osaka, Y.-s.; Masuko, T.; Kakehi, K. Anal. Biochem. 2007, 363, 245−257. Yamada, K.; Hyodo, S.; Matsuno, Y. K.; Kinoshita, M.; Maruyama, S. Z.; Osaka, Y. S.; Casal, E.; Lee, Y. C.; Kakehi, K. Anal. Biochem. 2007, 371, 52−61). The method allows rapid release of O-glycans as the reducing form within 10 min. In the present study, we connected the device to a MALDI-TOF MS spotter and achieved routine analysis of O-glycans in biological samples for clinical use after in situ derivatization of the released O-glycans with phenylhydrazine. We applied the method to the analysis of O-glycans expressed on MKN45 cells derived from human stomach cancer cells and found that MKN45 cells expressed characteristic trisialo-polylactosamine-type glycans as reported previously (Yamada, K.; Kinoshita, M.; Hayakawa, T.; Nakaya, S.; Kakehi, K. J. Proteome Res. 2009, 8, 521−537). We also applied the method to the analysis of O-glycans in serum samples. The present technique is the first attempt to use MS measurement for routine clinical diagnostic works.