The elucidation of key influence factors for electrostatic adsorption is very important to control protein nonspecific adsorption on modified surfaces. In this study, real time surface plasmon resonance technique is u...The elucidation of key influence factors for electrostatic adsorption is very important to control protein nonspecific adsorption on modified surfaces. In this study, real time surface plasmon resonance technique is used to characterize the electrostatic adsorption of two proteins (mouse IgG and protein A) on carboxymethyldextran modified surface. The results show that protein solution pH and ionic strength are key influence factors for efficient electrostatic adsorption. The influence of protein solution pH on the amount of electrostatic adsorption depends on the type of the charge and the charge density of both protein and modified matrix on the surface. The electrostatic adsorption process involves a competition between the positively charged protein and other positively charged species in the buffer solution. A decrease of ionic strength leads to an increasing electrostatic adsorption. The kinetic adsorption constants of protein A at different pH values were also calculated and compared.展开更多
人凝血因子7a(FVIIa)是外源性凝血途径的核心因子,检测参与凝血过程的FVIIa及针对FVIIa开发安全有效和经济的凝血剂和抗凝血剂是具有重要的意义。本研究基于指数富集配体系统进化技术(Systematic evolution of ligands by exponential e...人凝血因子7a(FVIIa)是外源性凝血途径的核心因子,检测参与凝血过程的FVIIa及针对FVIIa开发安全有效和经济的凝血剂和抗凝血剂是具有重要的意义。本研究基于指数富集配体系统进化技术(Systematic evolution of ligands by exponential enrichment,SELEX),采用酸纤维素膜法,通过11轮筛选成功分离出FVIIa的DNA适配体,并模拟了其二级结构。此适配体与FVIIa蛋白结合的表观解离常数为102nmol/L。同时,基于所获得的高亲和力适配体,建立了基于适配体的荧光检测方法。本方法简单高效,对FVIIa蛋白的定量检测范围达到30~80nmol/L,为人凝血因子7a的检测提供了一条新途径。非变性聚丙烯酰胺凝胶电泳和适配体的荧光检测方法结果表明,此适配体具有良好的选择性。展开更多
文摘The elucidation of key influence factors for electrostatic adsorption is very important to control protein nonspecific adsorption on modified surfaces. In this study, real time surface plasmon resonance technique is used to characterize the electrostatic adsorption of two proteins (mouse IgG and protein A) on carboxymethyldextran modified surface. The results show that protein solution pH and ionic strength are key influence factors for efficient electrostatic adsorption. The influence of protein solution pH on the amount of electrostatic adsorption depends on the type of the charge and the charge density of both protein and modified matrix on the surface. The electrostatic adsorption process involves a competition between the positively charged protein and other positively charged species in the buffer solution. A decrease of ionic strength leads to an increasing electrostatic adsorption. The kinetic adsorption constants of protein A at different pH values were also calculated and compared.
文摘人凝血因子7a(FVIIa)是外源性凝血途径的核心因子,检测参与凝血过程的FVIIa及针对FVIIa开发安全有效和经济的凝血剂和抗凝血剂是具有重要的意义。本研究基于指数富集配体系统进化技术(Systematic evolution of ligands by exponential enrichment,SELEX),采用酸纤维素膜法,通过11轮筛选成功分离出FVIIa的DNA适配体,并模拟了其二级结构。此适配体与FVIIa蛋白结合的表观解离常数为102nmol/L。同时,基于所获得的高亲和力适配体,建立了基于适配体的荧光检测方法。本方法简单高效,对FVIIa蛋白的定量检测范围达到30~80nmol/L,为人凝血因子7a的检测提供了一条新途径。非变性聚丙烯酰胺凝胶电泳和适配体的荧光检测方法结果表明,此适配体具有良好的选择性。
