Department of Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, PR China.
Biosens Bioelectron. 2013 Nov 15;49:20-4. doi: 10.1016/j.bios.2013.04.038. Epub 2013 May 3.
Herein, we report a simple, sensitive label-free colorimetric assay of trypsin based on silver nanoparticles (AgNPs) aggregation. Generally, a specially designed short peptide chain acts as both the stabilizer of AgNPs and the substrate of trypsin. In the presence of trypsin, the negatively charged part of peptides will be hydrolyzed, leaving the positively charged dipeptide capped on the surface of AgNPs. The electrostatic property alteration then leads to the AgNPs' aggregation in certain salt condition. The solution color may change correspondingly due to the localized surface plasmon resonance, which can be monitored by naked eye and UV-vis spectrophotometry. This novel AgNPs-based colorimetric method for quantitative determination of trypsin has a linear detection range from 2.5 to 200 ng mL(-1) and a rather low detection limit down to 2 ng mL(-1). The determination of trypsin can also be realized in complex biological fluids by the proposed method, demonstrating its great potential utility in the clinical applications in the future.
本文报道了一种基于银纳米粒子(AgNPs)聚集的简单、灵敏的无标记比色法测定胰蛋白酶。通常,一种特殊设计的短肽链既可以作为 AgNPs 的稳定剂,也可以作为胰蛋白酶的底物。在胰蛋白酶存在的情况下,肽的带负电荷部分会被水解,留下带正电荷的二肽帽在 AgNPs 的表面。这种静电性质的改变导致在一定盐条件下 AgNPs 的聚集。由于局域表面等离子体共振,溶液颜色会相应地发生变化,这可以通过肉眼和紫外可见分光光度法进行监测。这种基于新型 AgNPs 的比色法可用于定量测定胰蛋白酶,其线性检测范围为 2.5 至 200 ng mL(-1),检测限低至 2 ng mL(-1)。该方法还可以在复杂的生物流体中实现胰蛋白酶的测定,表明其在未来临床应用中有很大的潜力。