A label-free optical technique for detecting small molecule interactions
A label-free optical technique for detecting small molecule interactions
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DOI：
10.1016/s0956-5663(02)00077-5
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发表时间：
2002-09-01
影响因子：
12.6
通讯作者：
Cunningham, B
中科院分区：
文献类型：
--
作者：
Lin, B;Qiu, J;Cunningham, B
提出了一种利用比色共振光栅作为表面结合平台的分子相互作用的无标记检测新方法。当用白光照明时，这种光栅被设计成只反射单一波长。当分子附着在表面时，由于耦合到光栅中的光的光路的改变，反射波长(颜色)移动。通过将受体分子连接到光栅表面，可以在不使用任何类型的荧光探针或放射性标签的情况下检测到互补结合分子。该检测技术能够检测小分子与传感器表面的受体分子或传感器周围溶液中的酶相互作用时的添加和移除。提出了两种检测小分子与生物传感器相互作用的方法。首先，用亲和素受体层检测244Da的生物素结合。其次，进行了从固定化底物上裂解136Da对硝基苯胺(PNA)基团的酶活性测定。由于传感器结构可以嵌入到微滴定板的塑料表面或微阵列载玻片的玻璃表面，预计这项技术将在并行测量大量生物分子相互作用的应用中最有用，特别是当分子标签将改变或抑制所研究分子的功能时。筛选具有蛋白质靶标的药物化合物文库，以及蛋白质组学中蛋白质相互作用的微阵列筛选是需要这种方法所提供的灵敏度和吞吐量的应用的例子。(C)2002 Elsevier Science B.V.保留所有权利。
A novel approach for the label-free detection of molecular interactions is presented in which a colorimetric resonant grating is used as a surface binding platform. The grating, when illuminated with white light, is designed to reflect only a single wavelength. When molecules are attached to the surface, the reflected wavelength (color) is shifted due to the change of the optical path of light that is coupled into the grating. By linking receptor molecules to the grating surface, complementary binding molecules can be detected without the use of any kind of fluorescent probe or radioactive label. The detection technique is capable of detecting the addition and removal of small molecules as they interact with receptor molecules on the sensor surface or enzymes in the solution surrounding the sensor. Two assays are presented to exemplify the detection of small molecule interactions with the biosensor. First, an avidin receptor layer is used to detect 244 Da biotin binding. Second, a protease assay is performed in which a 136 Da p-nitroanilide (pNA) moeity is cleaved from an immobilized substrate. Because the sensor structure can be embedded in the plastic surfaces of microtiter plates or the glass surfaces of microarray slides, it is expected that this technology will be most useful in applications where large numbers of biomolecular interactions are measured in parallel, particularly when molecular labels will alter or inhibit the functionality of the molecules under study. Screening of pharmaceutical compound libraries with protein targets, and microarray screening of protein protein interactions for proteomics are examples of applications that require the sensitivity and throughput afforded by this approach. (C) 2002 Elsevier Science B.V. All rights reserved.