Competitive fluorescence anisotropy/polarization assay for ATP using aptamer as affinity ligand and dye-labeled ATP as fluorescence tracer
文献类型：期刊论文
| 作者 | Li, Yapiao; Sun, Linlin; Zhao, Qiang |
| 刊名 | TALANTA
 |
| 出版日期 | 2017-05-02 |
| 卷号 | 174期号:0页码:7-13 |
| 关键词 | Fluorescence Anisotropy Fluorescence Polarization Signal Amplification Adenosine Triphosphate Competitive Assay Fluorescence Tracer |
| 英文摘要 | We developed an aptamer-based competitive fluorescence anisotropy (FA)/fluorescence polarization (FP) assay for adenosine triphosphate (ATP). Different from the traditional fluorescence polarization immunoassays for small molecules, here DNA aptamer against ATP was used as affinity ligand, and tetramethylrhodamine (TMR) labeled ATP served as fluorescent tracer. The binding between TMR-labeled ATP and aptamer gave large FA due to molecular volume increase and restricted rotation of the dye -labeled ATP. When ATP was added in solution, ATP competitively displaced the TMR-labeled ATP from aptamer affinity complex, causing decrease of FA of TMR-labeled ATP. The buffer containing MgCl2 and incubation at low temperature were preferred for large FA change in the FA assay. The FA change was further enhanced in this competitive FA assay by increasing the molecular weight of aptamer through extension of aptamer sequences or conjugating streptavidin protein on aptamer. This method allowed for the detection of ATP in the range from 0.5 mu M to 1 mM, generating the maximum FA change about0.187 (corresponding maximum FP change about 0.242). The detection of ATP spiked in diluted urine or serum sample was achieved, showing capability for analysis in complex sample matrix. This assay also enabled the detection of the analogues of ATP, e.g. adenosine, adenosine monophosphate (AMP), and adenosine diphosphate (ADP) with similar sensitivity. This aptamer-based competitive FA assay takes advantages of aptamer in ease of synthesis, good thermal stability, and facile modulating the molecular mass of aptamer. |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/39276]  |
| 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 | Li, Yapiao,Sun, Linlin,Zhao, Qiang. Competitive fluorescence anisotropy/polarization assay for ATP using aptamer as affinity ligand and dye-labeled ATP as fluorescence tracer[J]. TALANTA,2017,174(0):7-13. |
| APA | Li, Yapiao,Sun, Linlin,&Zhao, Qiang.(2017).Competitive fluorescence anisotropy/polarization assay for ATP using aptamer as affinity ligand and dye-labeled ATP as fluorescence tracer.TALANTA,174(0),7-13. |
| MLA | Li, Yapiao,et al."Competitive fluorescence anisotropy/polarization assay for ATP using aptamer as affinity ligand and dye-labeled ATP as fluorescence tracer".TALANTA 174.0(2017):7-13. |
入库方式： OAI收割
来源：生态环境研究中心
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