Department of Chemical and Biomolecular Engineering, 4 Engineering Drive 4, National University of Singapore, 117576 Singapore.
Langmuir. 2010 Jun 15;26(12):10025-30. doi: 10.1021/la100139p.
An assay triggered by recognition-induced charge switching is developed for protein detection and quantification. Aptamer-functionalized silica nanoparticles (NPs) have been synthesized to capture lysozyme, resulting in an alternation of the surface charge from negative to partially positive. The binding event is then translated and monitored by the fluorescence signal of a highly fluorescent anionic poly(fluorene-alt-vinylene) (PFVSO(3)), which "stains" on protein/aptamer-NP complexes via electrostatic interaction. Blue-greenish fluorescence of PFVSO(3) is observed in the presence of lysozyme by the naked eye, while no fluorescence is obtained for NPs upon treatment with a mixture of foreign proteins. A linear relationship between NP fluorescence and lysozyme is observed in the concentration range of 0-22.5 microg/mL, which gives a limit of detection as approximately 0.36 microg/mL. This work demonstrates a convenient label-free conjugated polyelectrolyte (CPE)-based protein detection with high specificity and sensitivity, which has potential applications in medical diagnosis.
一种基于识别诱导电荷转换的分析方法被开发用于蛋白质的检测和定量。合成了适配体功能化的硅纳米粒子(NPs)以捕获溶菌酶,导致表面电荷从负到部分正的转变。然后,通过高度荧光的阴离子聚(芴-alt-乙烯基)(PFVSO(3))的荧光信号来转化和监测结合事件,该荧光信号通过静电相互作用“染色”在蛋白质/适配体-NP 复合物上。在存在溶菌酶的情况下,肉眼可以观察到 PFVSO(3)的蓝绿色荧光,而用混合的外源蛋白处理 NPs 时则没有荧光。在 0-22.5μg/mL 的浓度范围内观察到 NP 荧光与溶菌酶之间存在线性关系,其检测限约为 0.36μg/mL。这项工作展示了一种方便的无标记共轭聚合物电解质(CPE)基蛋白质检测方法,具有高特异性和灵敏度,在医学诊断中有潜在的应用。