A lectin array-based methodology for the analysis of protein glycosylation

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• 作者：Rakefet Rosenfeld ; Haim Bangio ; Gerrit J. Gerwig ; Revital Rosenberg ; Ronny Aloni ; Yossi Cohen ; Yehudit Amor ; Inbar Plaschkes ; Johannis P. Kamerling ; Ruth Ben-Yakar Maya
• 关键词：Glycosylation ; Glycoanalysis ; Lectins ; Protein arrays ; Proteomics ; Glycomics
• 刊名：Journal of Biochemical and Biophysical Methods
• 出版年：2007
• 出版时间：10 April 2007
• 年：2007
• 卷：70
• 期：3
• 页码：415-426
• 全文大小：1109 K

文摘

Glycosylation is the most versatile and one of the most abundant protein modifications. It has a structural role as well as diverse functional roles in many specific biological functions, including cancer development, viral and bacterial infections, and autoimmunity. The diverse roles of glycosylation in biological processes are rapidly growing areas of research, however, Glycobiology research is limited by the lack of a technology for rapid analysis of glycan composition of glycoproteins. Currently used methods for glycoanalysis are complex, typically requiring high levels of expertise and days to provide answers, and are not readily available to all researcher.

We have developed a lectin array-based method, Qproteome™ GlycoArray kits, for rapid analysis of glycosylation profiles of glycoproteins. Glycoanalysis is performed on intact glycoproteins, requiring only 4–6 h for most analysis types. The method, demonstrated in this manuscript by several examples, is based on binding of an intact glycoprotein to the arrayed lectins, resulting in a characteristic fingerprint that is highly sensitive to changes in the protein's glycan composition. The large number of lectins, each with its specific recognition pattern, ensures high sensitivity to changes in the glycosylation pattern. A set of proprietary algorithms automatically interpret the fingerprint signals to provide a comprehensive glycan profile output.