College of Chemistry and Chemical Engineering, University of South China, Hengyang, Hunan 421001, China.
Biosens Bioelectron. 2012 Apr 15;34(1):106-11. doi: 10.1016/j.bios.2012.01.025. Epub 2012 Jan 28.
In this study, we report a double-receptor sandwich supramolecule sensing method for the determination of adenosine triphosphate (ATP). One receptor is a uranyl-salophen complex which can bind the triphosphate group in ATP selectively, and another is an anti-adenosine aptamer which is a single-stranded oligonucleotide and can recognize the adenosine group in ATP specifically. The uranyl-salophen complex was immobilized on the surface of amino-silica gel particles and used as the solid phase receptor of ATP. The anti-adenosine aptamer was labeled with a fluorescent group and used as the labeled receptor of ATP. In the procedure of ATP detection, ATP was first combined with the solid phase receptor and then conjugated with the labeled receptor to form a sandwich-type supramolecule. The conditions of fabricating solid phase receptor and the influence of manifold variables on the determination were studied. The experimental results demonstrate that the method has a number of advantages such as high selectivity, high sensitivity, good stability and low cost. Under optimal conditions, the linear range for detection of ATP is 0.2-5.0 nmol/mL with a detection limit of 0.037 nmol/mL. The proposed method was successfully applied for the determination of ATP in real samples with the recoveries of 96.8-103.3%.
在本研究中，我们报告了一种用于测定三磷酸腺苷（ATP）的双受体夹心超分子传感方法。一个受体是铀酰-salophen 配合物，它可以选择性地结合 ATP 中的三磷酸基团，另一个受体是抗腺苷适体，它是一种单链寡核苷酸，能够特异性地识别 ATP 中的腺苷基团。铀酰-salophen 配合物被固定在氨基硅烷凝胶颗粒的表面上，用作 ATP 的固相受体。抗腺苷适体被标记上荧光基团，用作 ATP 的标记受体。在 ATP 检测过程中，ATP 首先与固相受体结合，然后与标记受体结合，形成夹心型超分子。研究了制备固相受体的条件和多种变量对测定的影响。实验结果表明，该方法具有高选择性、高灵敏度、良好的稳定性和低成本等优点。在最佳条件下，检测 ATP 的线性范围为 0.2-5.0 nmol/mL，检测限为 0.037 nmol/mL。该方法成功地应用于实际样品中 ATP 的测定，回收率为 96.8-103.3%。
