A General Chemiluminescence Strategy for Measuring Aptamer鈥揟arget Binding and Target Concentration

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• 作者：Shiyuan Li ; Duyu Chen ; Qingtong Zhou ; Wei Wang ; Lingfeng Gao ; Jie Jiang ; Haojun Liang ; Yangzhong Liu ; Gaolin Liang ; Hua Cui
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：June 3, 2014
• 年：2014
• 卷：86
• 期：11
• 页码：5559-5566
• 全文大小：473K
• 年卷期：v.86,no.11(June 3, 2014)
• ISSN：1520-6882

文摘

Although much effort has been made for studies on aptamer鈥搕arget interactions due to promising applications of aptamers in biomedical and analytical fields, measurement of the aptamer鈥搕arget binding constant and binding site still remains challenging. Herein, we report a sensitive label-free chemiluminescence (CL) strategy to determine the target concentration and, more importantly, to measure the target鈥揳ptamer binding constant and binding site. This approach is suitable for multiple types of targets, including small molecules, peptides, and proteins that can enhance the CL initiated by N-(aminobutyl)-N-ethylisoluminol functionalized gold colloids, making the present method a general platform to investigate aptamer鈥搕arget interactions. This approach can achieve extremely high sensitivity with nanogram samples for measuring the target鈥揳ptamer binding constant. And the measurement could be rapidly performed using a simple and low-cost CL system. It provides an effective tool for studying the binding of biologically important molecules to nucleic acids and the selection of aptamers. Besides, we have also discovered that the 14-mer aptamer fragment itself split from the ATP-binding aptamer could selectively capture ATP. The binding constant, site, and conformation between ATP and the 14-mer aptamer fragment were obtained using such a novel CL strategy and molecular dynamic simulation.