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碲化镉量子点与溶菌酶结合DNA的非共价共轭用于复杂生物样品中溶菌酶的荧光传感

Non-covalent conjugation of CdTe QDs with lysozyme binding DNA for fluorescent sensing of lysozyme in complex biological sample.

作者信息

Li Shujia, Gao Zhidan, Shao Na

机构信息

College of Chemistry, Beijing Normal University, Beijing 100875, PR China.

College of Chemistry, Beijing Normal University, Beijing 100875, PR China.

出版信息

Talanta. 2014 Nov;129:86-92. doi: 10.1016/j.talanta.2014.04.062. Epub 2014 May 2.

Abstract

Water-soluble cysteamine (CA) capped CdTe quantum dots (QDs) conjugated with lysozyme binding DNA (LBD) was constructed for luminescent sensing of lysozyme by forming a ternary self-assembly complex. Addition of negatively charged lysozyme binding DNA to the positively charged CA capped CdTe QDs buffer solution (Tris-HCl pH 7.4) could lead to the formation of QDs-LBD complex through electrostatic interactions. Once lysozyme was introduced into the CdTe QDs-LBD system, it could bind specifically with the QDs-LBD complex, resulting in fluorescence emission enhancement of the QDs due to the surface inert of QDs. At a given amount of LBD and CdTe QDs (LBD: QDs=2: 1), the fluorescence intensity enhancement of QDs was linear with lysozyme concentration over the range of 8.9-71.2 nM, with a detection limit of 4.3 nM. Due to the specific binding of LBD with lysozyme, this approach displayed high selectivity for lysozyme recognition. The sensing mechanism was confirmed by DLS and zeta potential measurement, and agarose gel electrophoresis experiment. Furthermore, the proposed CA-capped CdTe QDs-LBD sensor was applied to lysozyme detection in mouse serum and human morning urine samples, which showed high sensitivity and selectivity in the complex biological sample.

摘要

构建了与溶菌酶结合DNA(LBD)共轭的水溶性半胱胺(CA)包覆的碲化镉量子点(QDs),通过形成三元自组装复合物用于溶菌酶的发光传感。向带正电的CA包覆的CdTe量子点缓冲溶液(Tris-HCl pH 7.4)中加入带负电的溶菌酶结合DNA,可通过静电相互作用形成量子点-LBD复合物。一旦将溶菌酶引入CdTe量子点-LBD系统,它就能与量子点-LBD复合物特异性结合,由于量子点的表面惰性,导致量子点的荧光发射增强。在给定的LBD和CdTe量子点量(LBD:量子点 = 2:1)下,量子点的荧光强度增强与溶菌酶浓度在8.9 - 71.2 nM范围内呈线性关系,检测限为4.3 nM。由于LBD与溶菌酶的特异性结合,该方法对溶菌酶识别具有高选择性。通过动态光散射(DLS)、zeta电位测量和琼脂糖凝胶电泳实验证实了传感机制。此外,所提出的CA包覆的CdTe量子点-LBD传感器应用于小鼠血清和人晨尿样品中的溶菌酶检测,在复杂生物样品中显示出高灵敏度和选择性。

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