Chuang Yen-Jun, Zhou Xichun, Pan Zhengwei, Turchi Craig
Faculty of Engineering, University of Georgia, Athens, GA 30602, USA.
Biochem Biophys Res Commun. 2009 Nov 6;389(1):22-7. doi: 10.1016/j.bbrc.2009.08.079. Epub 2009 Aug 19.
Carbohydrate functionalized nanoparticles, i.e., the glyconanoparticles, have wide application ranging from studies of carbohydrate-protein interactions, in vivo cell imaging, biolabeling, etc. Currently reported methods for preparation of glyconanoparticles require multi-step modifications of carbohydrates moieties to conjugate to nanoparticle surface. However, the required synthetic manipulations are difficult and time consuming. We report herewith a simple and versatile method for preparing glyconanoparticles. This method is based on the utilization of clean and convenient microwave irradiation energy for one-step, site-specific conjugation of unmodified carbohydrates onto hydrazide-functionalized Au nanoparticles. A colorimetric assay that utilizes the ensemble of gold glyconanoparticles and Concanavalin A (ConA) was also presented. This feasible assay system was developed to analyze multivalent interactions and to determine the dissociation constant (K(d)) for five kind of Au glyconanoparticles with lectin. Surface plasmon changes of the Au glyconanoparticles as a function of lectin-carbohydrate interactions were measured and the dissociation constants were determined based on non-linear curve fitting. The strength of the interaction of carbohydrates with ConA was found to be as follows: maltose>mannose>glucose>lactose>MAN5.
碳水化合物功能化纳米粒子,即糖纳米粒子,在碳水化合物 - 蛋白质相互作用研究、体内细胞成像、生物标记等方面有着广泛应用。目前报道的制备糖纳米粒子的方法需要对碳水化合物部分进行多步修饰以连接到纳米粒子表面。然而,所需的合成操作困难且耗时。我们在此报告一种简单且通用的制备糖纳米粒子的方法。该方法基于利用清洁便捷的微波辐射能量,将未修饰的碳水化合物一步、位点特异性地连接到酰肼功能化的金纳米粒子上。还介绍了一种利用金糖纳米粒子和伴刀豆球蛋白A(ConA)的比色测定法。开发这个可行的测定系统是为了分析多价相互作用,并确定五种金糖纳米粒子与凝集素的解离常数(K(d))。测量了金糖纳米粒子的表面等离子体变化与凝集素 - 碳水化合物相互作用的关系,并基于非线性曲线拟合确定了解离常数。发现碳水化合物与ConA的相互作用强度如下:麦芽糖>甘露糖>葡萄糖>乳糖>MAN5。