Quantitative Assessment of the Multivalent Protein鈥揅arbohydrate Interactions on Silicon

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• 作者：Jie Yang ; Jean-No毛l Chazalviel ; Aloysius Siriwardena ; Rabah Boukherroub ; Fran莽ois Ozanam ; Sabine Szunerits ; Anne Chantal Gouget-Laemmel
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：October 21, 2014
• 年：2014
• 卷：86
• 期：20
• 页码：10340-10349
• 全文大小：714K
• ISSN：1520-6882

文摘

A key challenge in the development of glycan arrays is that the sensing interface be fabricated reliably so as to ensure the sensitive and accurate analysis of the protein鈥揷arbohydrate interaction of interest, reproducibly. These goals are complicated in the case of glycan arrays as surface sugar density can influence dramatically the strength and mode of interaction of the sugar ligand at any interface with lectin partners. In this Article, we describe the preparation of carboxydecyl-terminated crystalline silicon (111) surfaces onto which are grafted either mannosyl moieties or a mixture of mannose and spacer alcohol molecules to provide 鈥渄iluted鈥?surfaces. The fabrication of the silicon surfaces was achieved efficiently through a strategy implicating a 鈥渃lick鈥?coupling step. The interactions of these newly fabricated glycan interfaces with the lectin, Lens culinaris, have been characterized using quantitative infrared (IR) spectroscopy in the attenuated total geometry (ATR). The density of mannose probes and lectin targets was precisely determined for the first time by the aid of special IR calibration experiments, thus allowing for the interpretation of the distribution of mannose and its multivalent binding with lectins. These experimental findings were accounted for by numerical simulations of lectin adsorption.