Gold nanoparticles-based protease assay
Gold nanoparticles-based protease assay
复制标题
DOI：
10.1073/pnas.0509372103
复制
发表时间：
2006-03-14
影响因子：
11.1
通讯作者：
Scrimin, P
中科院分区：
文献类型：
--
作者：
Guarise, C;Pasquato, L;Scrimin, P
我们在这里描述了一个简单的测定，允许蛋白酶的视觉检测。该方法利用金胶体等离子体带的高摩尔吸光系数，并基于用二硫醇处理时其溶液的颜色变化。我们使用的C-和N-末端半胱氨酰衍生物的肽底物利用其选择性识别和切割的特定蛋白酶。与天然肽相反，裂解的肽不能诱导纳米颗粒聚集;因此，溶液的颜色不会改变。报告了两种蛋白酶的检测：凝血酶（参与血液凝固和血栓形成）和致死因子（炭疽杆菌产生的毒素的酶组分）。这种基于纳米颗粒的测定的灵敏度在低纳摩尔范围内。
We describe here a simple assay that allows the visual detection of a protease. The method takes advantage of the high molar absorptivity of the plasmon band of gold colloids and is based on the color change of their solution when treated with dithiols. We used C- and N-terminal cysteinyl derivatives of a peptide substrate exploiting its selective recognition and cleavage by a specific protease. Contrary to the native ones, cleaved peptides are unable to induce nanoparticles aggregation; hence, the color of the solution does not change. The detection of two proteases is reported: thrombin (involved in blood coagulation and thrombosis) and lethal factor (an enzyme component of the toxin produced by Bacillus anthracis). The sensitivity of this nanoparticle-based assay is in the low nanomolar range.