Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, People's Republic of China.
Analyst. 2013 Mar 7;138(5):1475-82. doi: 10.1039/c2an36269d.
A simple, colorimetric 'turn on' sensor for ultrasensitive detection of thrombin has been developed using fibrinogen-modified gold nanoparticles (Fib-Au NPs). The assay was based on the thrombin-fibrinogen interaction which is part of the physiological process of blood clotting. The fibrinogen was immobilized on the surface of 96-well plate offering reactive N-oxysuccinimide esters (referred to as NOS group) surface. Introducing thrombin and Fib-Au NPs into the fibrinogen-bound 96-well plate induced the immobilization of Fib-Au NPs on the surface of 96-well plate through the thrombin mediated conversion of soluble fibrinogen to insoluble cross-linked fibrin. Such process could be detected visually post HAuCl(4)-NH(2)OH redox reaction catalyzed by the Au NPs. The parameters governing the performance of the assay have been optimized. The detection limit was 3.2 fM, corresponding to 0.16 amol thrombin in 50 μL of sample. Other proteins, such as bovine serum albumin (BSA), pepsin, trypsin, hemoglobin, lysozyme, and cytochrome c did not show interference with the assay of thrombin. In addition, the work demonstrates the feasibility of thrombin detection in a complex matrix, showing potential for rapid medical diagnostics.
一种简单的、比色的“开启”传感器，用于超灵敏检测凝血酶，是使用纤维蛋白原修饰的金纳米粒子（Fib-Au NPs）开发的。该测定法基于凝血酶-纤维蛋白原相互作用，这是血液凝固生理过程的一部分。纤维蛋白原被固定在 96 孔板的表面，提供反应性 N-羟基琥珀酰亚胺酯（称为 NOS 基团）表面。将凝血酶和 Fib-Au NPs 引入纤维蛋白原结合的 96 孔板中，通过凝血酶介导的可溶性纤维蛋白原转化为不溶性交联纤维蛋白，诱导 Fib-Au NPs 在 96 孔板表面的固定。这种过程可以通过 Au NPs 催化的 HAuCl(4)-NH(2)OH 氧化还原反应后进行目视检测。已经优化了测定法性能的参数。检测限为 3.2 fM，相当于 50 μL 样品中 0.16 amol 的凝血酶。其他蛋白质，如牛血清白蛋白（BSA）、胃蛋白酶、胰蛋白酶、血红蛋白、溶菌酶和细胞色素 c 不会干扰凝血酶的测定。此外，该工作证明了在复杂基质中检测凝血酶的可行性，显示出快速医疗诊断的潜力。
