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• Yagi, H
• Ohno, E
• Kondo, S
• Yoshida, A
• Kato, K
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• Yagi, H
• Ohno, E
• Kondo, S
• Yoshida, A
• Kato, K
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• Yagi, H
• Ohno, E
• Kondo, S
• Yoshida, A
• Kato, K

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Biomolecules 2011, 1(1), 48-62; doi:10.3390/biom1010048

Article

Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides

Hirokazu Yagi ¹ , Erina Ohno ¹ , Sachiko Kondo ^1,4 , Atsuhiro Yoshida ²  and Koichi Kato ^1,3,4,5,*

¹ Graduate School of Pharmaceutical Science, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan ² Graduate School of Medical Sciences and Medical School, Nagoya City University, Kawasumi-1, Mizuho-cho Mizuho-ku, Nagoya 467-8601, Japan ³ Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, 5-1 Higashiyama Myodaiji, Okazaki 444-8787, Japan ⁴ GLYENCE Co., Ltd., 2-22-8 Chikusa, Chikusa-ku, Nagoya 464-0858, Japan ⁵ The Glycoscience Institute, Ochanomizu University, 2-1-1 Ohtsuka, Bunkyo-ku, Tokyo 112-8610, Japan

* Author to whom correspondence should be addressed.

Received: 29 August 2011; in revised form: 30 November 2011 / Accepted: 5 December 2011 / Published: 14 December 2011

(This article belongs to the Special Issue Feature Papers)

Download PDF Full-Text [454 KB, uploaded 14 December 2011 16:25 CET]

Abstract: Glycosylation improves the solubility and stability of proteins, contributes to the structural integrity of protein functional sites, and mediates biomolecular recognition events involved in cell-cell communications and viral infections. The first step toward understanding the molecular mechanisms underlying these carbohydrate functionalities is a detailed characterization of glycan structures. Recently developed glycomic approaches have enabled comprehensive analyses of N-glycosylation profiles in a quantitative manner. However, there are only a few reports describing detailed O-glycosylation profiles primarily because of the lack of a widespread standard method to identify O-glycan structures. Here, we developed an HPLC mapping method for detailed identification of O-glycans including neutral, sialylated, and sulfated oligosaccharides. Furthermore, using this method, we were able to quantitatively identify isomeric products from an in vitro reaction catalyzed by N-acetylglucosamine-6O-sulfotransferases and obtain O-glycosylation profiles of serum IgA as a model glycoprotein.

Keywords: O-glycans; HPLC map; glycosylation profiling; sulfated oligosaccharide; IgA

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