一种糖生物传感芯片制备的新方法  

A Novel Method of Immobilization of 911 for Surface Plasmon Resonance Biosensor

作　　者：李福川[1] 戚欣[1] 耿美玉[1] 

机构地区：[1]中国海洋大学海洋药物与食品研究所,青岛266003

出　　处：《分析化学》2003年第3期266-270,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金资助项目 (No .30 0 70 694;30 130 2 0 0 )

摘　　要：建立了一种用于SPR生物传感器分析的糖生物传感芯片制备新方法。在己二胺大量过量的情况下 ,海洋硫酸多糖 91 1还原末端的半缩醛基与己二胺的一个氨基进行还原胺化反应 ,引入一个伯氨基 ,该伯氨基进一步与sulfo NHS biotin反应 ,使 91 1的还原末端生物素化 ,通过预偶联到CM5芯片上的链酶抗生物素抗体 ,以俘获法将生物素化的 91 1固定到芯片表面。该方法避免已有固定方法对糖内部结构的破坏 ,减少了非特异性耦联 ,能更好的反映糖类与其它分子的相互作用特性 ,且操作简单 ,适应于多数具有还原末端的糖类。91 1经该方法固定后 ,利用SPR生物传感器 ,研究了其与HIVgp1 2 0蛋白V3区的相互作用动力学 ,初步证明两者之间存在强烈的相互作用 ,KA 与KD 分别为 :2 .2 5e5与 4 .4 4e -6。A method for the immobilization of polysaccharides to the biosensor chip surface is described. The reductive end of 911 was introduced for primary amine by reductive amination reaction with 1,6-hexanediamine and was biotinylated further via the reaction of the primary amine with sulfo-NHS-biotin. The biotinylated 911 via reductive end was immobilized on the surface of biosensor chip CM5 by streptavidin-biotin coupling. Comparing with being reported methods, this method avoided impairing the internal structure of polysacccharide as well as being simple and applicable for the immobilization of many reductive polysaccharides and oligosaccharides. Surface plasmon resonance (SPR) was used to monitor the interaction of marine sulfated polysaccharide-911 with V3 region of HIV-1 MN gp120, empolying 911 immobilized on a sensor chip surface. The interaction of 911 with V3 region was discussed. The dissociation constants were estimated to be approximately. Thus, V3 region appeared with high affinity to 911.

关 键 词：生物传感芯片 制备 糖 表面等离子体共振 固定 生物传感器 分析 抗爱滋病药物 海洋硫酸多糖 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置]