多色彩酶联免疫吸附测定

In the presence of different concentrations of the target, the traditional enzyme-linked immunosorbent assay (ELISA) typically produces only color depth changes. But the naked eye is more sensitive to color changes than the depth change of the same color, so the conventional ELISA can only be applied in the qualitative analysis of the targets and special instruments are needed for quantitative measurement. TMB2+, the chromogenic substrate of conventional ELISA, which can react with the gold nanorods and causing the changing of aspect ratios of the nanorods, which results in the series changing of the color of the system, based on which, a multi-color ELISA can be developed for the sensitive visual detection of the targets..The study intends to focus on the following aspects: (1) Study about the correspondence between the aspect ratio of the gold nanorods and the display color of the solution; (2) Systematic study about the mechanism of the reaction between TMB2+ and gold nanorods and deduce the quantitative relationship between the concentration of TMB2+ and the aspect ratio of the gold nanorods;(3) Systematic study about the relationship between the HRP concentration and the degree of the oxidation of the gold nanorods;(4)Design some multi-color ELISA kits and realize visual quantitative determination for different targets(specially for tumor markers).

在不同浓度目标物存在条件下，传统的酶联免疫分析法（ELISA）通常只产生颜色深浅的变化。然而人类肉眼对于同一种颜色的深浅变化并不敏感，因此传统ELISA通常只能通过肉眼对目标物进行定性分析，若欲进行定量测定通常需要借助一些专门的仪器才能实现。本项目根据传统ELISA中的底物四甲基联苯胺（TMB2+）能够与金纳米棒反应，从而使其长径比发生变化，进而引起溶液发生多种颜色变化的特点构建一种多色彩的ELISA新方法，从而达到对目标物进行目测定量的目的。.本项目拟重点研究以下几方面内容：(1)系统研究金纳米棒长径比与溶液所显示颜色的对应关系；（2）详细研究TMB2+与金纳米棒的反应机理，从而最终得出TMB2+使金纳米棒变短的定量关系；（3）系统研究ELISA显色过程中的酶标物（HRP）浓度与金纳米棒氧化程度的关系；（4）开发多色ELISA试剂盒，实现对不同目标物（特别是肿瘤标志物）的可视化定量测定。

传统的酶联免疫分析法（ELISA）通常只产生单种颜色深浅的变化，利用肉眼无法进行定量分析。本课题根据金纳米球聚集状态的变化以及金纳米棒、金纳米锥的形貌变化引起的颜色变化，开发了一系列基于多种颜色变化的半定量（或定量）的肉眼可视化ELISA新方法。主要完成工作如下：（1）基于TMB2+能够有效刻蚀金纳米颗粒，构建了双色响应的plasmonic ELISA方法并用于灵敏地可视化检测前列腺特异性抗原；根据金纳米颗粒不同聚集态所呈现出的颜色的差异，结合信号放大技术和纳米金粒子的定向排列，构建了高灵敏检查miRNA的比色传感器；（2）根据金纳米棒长径比与溶液所显示颜色的对应变化关系，结合酶联免疫分析技术，构建了多色彩可视化生物传感器并应用于肿瘤标志物和食品安全等的分析；（3）利用金纳米双锥尖端的形貌的变化可以实现多色显色的特点，构建了多彩的免疫传感器并应用于疾病标志物等的肉眼可视化分析检测。.本项目执行期间共发表与本项目相关的SCI论文18篇，包括Anal. Chem. (4篇) 、Sens. Actuators, B (3篇)、 ACS Sens. (1篇)、Biosens. Bioelectron. (1篇)以及分析化学（1篇）等。本项目执行期间共获得授权发明专利3项。已培养博士2人，硕士5人。