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基于适体的荧光生物传感器,用于使用芘标记的分子信标检测钾离子。

An aptamer-based fluorescent biosensor for potassium ion detection using a pyrene-labeled molecular beacon.

机构信息

College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, People's Republic of China.

出版信息

Anal Biochem. 2010 May 1;400(1):99-102. doi: 10.1016/j.ab.2009.12.034. Epub 2010 Jan 6.

DOI:10.1016/j.ab.2009.12.034
PMID:20056100
Abstract

A novel and sensitive biosensor based on aptamer and pyrene-labeled fluorescent probes for the determination of K+ was developed. The aptamer was used as a molecular recognition element and a partially complementary oligonucleotide with the aptamer was labeled by pyrene moieties at both ends to transduce the binding event of K+ with aptamer. In the presence of K+, the complementary oligonucleotides were displaced from aptamers, which was accompanied by excimer fluorescence of pyrenes because the self-hairpin structure of the complementary oligonucleotide brought pyrene moieties into close proximity. However, it gave only monomer emission in the absence of K+. Under optimum conditions, the relative fluorescence intensity of pyrene was proportional to the concentration of K+ in the range of 6.0 x 10(-4) to 2.0 x 10(-2) M. A detection limit of 4.0 x 10(-4) M was achieved. Moreover, this method was able to detect K+ with high selectivity in the presence of Na+, NH4+, Mg2+, and Ca2+ ions of biological fluids. In brief, the assay may have great potential applications, especially in a biological environment because of its simplicity, sensitivity, and specificity.

摘要

基于适配体和芘标记荧光探针的新型灵敏生物传感器用于测定 K+。适配体用作分子识别元件,其部分互补寡核苷酸在两端用芘基团标记以转导 K+与适配体的结合事件。在存在 K+的情况下,互补寡核苷酸从适配体上置换出来,伴随着芘的激基缔合物荧光,因为互补寡核苷酸的自发夹结构使芘基团紧密接近。然而,在没有 K+的情况下,仅发出单体发射。在最佳条件下,芘的相对荧光强度与 6.0 x 10(-4) 至 2.0 x 10(-2) M 范围内的 K+浓度成正比。检测限达到 4.0 x 10(-4) M。此外,该方法在存在生物体液中的 Na+、NH4+、Mg2+和 Ca2+离子时能够具有高选择性地检测 K+。总之,由于其简单性、灵敏度和特异性,该测定方法可能具有很大的应用潜力,特别是在生物环境中。

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