Enhancing the Affinity of Anti-Human alpha-Thrombin 15-mer DNA Aptamer and Anti-Immunoglobulin E Aptamer by PolyT Extension

CORC > 生态环境研究中心 > 中国科学院生态环境研究中心 > 环境化学与生态毒理学国家重点实验室

	Enhancing the Affinity of Anti-Human alpha-Thrombin 15-mer DNA Aptamer and Anti-Immunoglobulin E Aptamer by PolyT Extension
	Wang, Hailin; Bai, Yunlong; Li, Yapiao; Zhang, Dapeng; Zhao, Qiang
刊名	ANALYTICAL CHEMISTRY
	2017-08-15
卷号	89 期号:17 页码:9467-9473
英文摘要	Aptamer affinity capillary electrophoresis laser-induced fluorescence (CE-LIF) for protein detection takes advantage of aptamers for their ease of synthesis and labeling, small size, and having many negative charges. Its success relies on the high binding affinity of aptamers. One 15-mer DNA aptamer (5'-GGT TGG TGT GGT TGG-3', Apt15) shows desirable specificity for human alpha-thrombin, an important enzyme with multiple functions in blood. However, Apt15 has weak binding affinity, and the use of Apt15 in affinity CELIF analysis remains challenging. Here we reported that extension of Apt15 at the 3'-end with a polyT tail having length of 18 T or longer significantly enhanced its affinity and enabled a well-isolated and stable peak for thrombin aptamer complex in affinity CE. It was likely that the improvement of binding affinity resulted from double binding, an additional interaction of the polyT tail with thrombin in addition to the Apt15 section binding to thrombin. With dye labeled Apt15 having a T-25 tail, we achieved detection of thrombin at concentrations as low as 0.1 nM by affinity CE-LIF. This aptamer probe specifically bound to human alpha-thrombin, showing negligible affinity for human beta- and gamma-thrombin, which are proteolyzed derivatives of human alpha alpha-thrombin and share similar structure. This strategy of adding a polyT extension also enhanced the binding affinity of anti-immunoglobulin E aptamer in CELIF analysis, showing that the affinity enhancement approach is not limited to the thrombin-binding aptamer and has potential for more applications in bioanalysis.
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/39306]
专题	生态环境研究中心_环境化学与生态毒理学国家重点实验室
推荐引用方式 GB/T 7714	Wang, Hailin,Bai, Yunlong,Li, Yapiao,et al. Enhancing the Affinity of Anti-Human alpha-Thrombin 15-mer DNA Aptamer and Anti-Immunoglobulin E Aptamer by PolyT Extension[J]. ANALYTICAL CHEMISTRY,2017,89(17):9467-9473.
APA	Wang, Hailin,Bai, Yunlong,Li, Yapiao,Zhang, Dapeng,&Zhao, Qiang.(2017).Enhancing the Affinity of Anti-Human alpha-Thrombin 15-mer DNA Aptamer and Anti-Immunoglobulin E Aptamer by PolyT Extension.ANALYTICAL CHEMISTRY,89(17),9467-9473.
MLA	Wang, Hailin,et al."Enhancing the Affinity of Anti-Human alpha-Thrombin 15-mer DNA Aptamer and Anti-Immunoglobulin E Aptamer by PolyT Extension".ANALYTICAL CHEMISTRY 89.17(2017):9467-9473.