Surface plasmon resonance: principles, methods and applications in biomedical sciences
Surface plasmon resonance: principles, methods and applications in biomedical sciences
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DOI：
10.1155/2003/372913
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发表时间：
2003-01-01
影响因子：
--
通讯作者：
Verhas, M
中科院分区：
文献类型：
--
作者：
Englebienne, P;Van Hoonacker, A;Verhas, M
表面等离子体共振 (SPR) 是当入射光束以特定角度照射金属表面（通常是金和银）时发生的一种现象。根据金属表面分子层的厚度，SPR 现象会导致反射光强度逐渐降低。生物医学应用利用 SPR 对金属表面附近介质的折射率的极高灵敏度，这使得精确测量分子在金属表面上的吸附及其最终与特定配体的相互作用成为可能。过去十年，SPR 在生物医学应用中的应用取得了巨大发展。该技术不仅应用于配体-受体相互作用动力学的实时测量和制药行业中先导化合物的筛选，而且还应用于多克隆抗体表征、表位作图、蛋白质构象研究和无标记免疫测定中DNA杂交、酶-底物相互作用的测量。传统的SPR应用于专门的生物传感仪器。这些仪器使用昂贵的传感器芯片，重复使用能力有限，并且需要复杂的化学方法来固定配体或蛋白质。我们的实验室已成功应用缓冲溶液中的胶体金颗粒进行SPR。与传统 SPR 相比，该应用具有许多优点。该支持物价格便宜，易于合成，并且可以通过电荷吸附的方式涂覆各种蛋白质或蛋白质-配体复合物。使用胶体金，可以在任何紫外-可见分光光度计中监测 SPR 现象。对于高通量应用，我们在自动化临床化学分析仪中采用了该技术。这种简单的技术可应用于蛋白质和小分析物的无标记定量免疫测定技术、蛋白质构象研究以及受体-配体相互作用的实时缔合-解离测量，用于高通量筛选和先导化合物优化。
Surface plasmon resonance (SPR) is a phenomenon occuring at metal surfaces (typically gold and silver) when an incident light beam strikes the surface at a particular angle. Depending on the thickness of a molecular layer at the metal surface, the SPR phenomenon results in a graded reduction in intensity of the reflected light. Biomedical applications take advantage of the exquisite sensitivity of SPR to the refractive index of the medium next to the metal surface, which makes it possible to measure accurately the adsorption of molecules on the metal surface and their eventual interactions with specific ligands. The last ten years have seen a tremendous development of SPR use in biomedical applications. The technique is applied not only to the measurement in real-time of the kinetics of ligand-receptor interactions and to the screening of lead compounds in the pharmaceutical industry, but also to the measurement of DNA hybridization, enzyme-substrate interactions, in polyclonal antibody characterization, epitope mapping, protein conformation studies and label-free immunoassays. Conventional SPR is applied in specialized biosensing instruments. These instruments use expensive sensor chips of limited reuse capacity and require complex chemistry for ligand or protein immobilization. Our laboratory has successfully applied SPR with colloidal gold particles in buffered solution. This application offers many advantages over conventional SPR. The support is cheap, easily synthesized, and can be coated with various proteins or protein-ligand complexes by charge adsorption. With colloidal gold, the SPR phenomenon can be monitored in any UV-vis spectrophotometer. For high-throughput applications, we have adapted the technology in an automated clinical chemistry analyzer. This simple technology finds application in label-free quantitative immunoassay techniques for proteins and small analytes, in conformational studies with proteins as well as in the real-time association-dissociation measurements of receptor-ligand interactions, for high-throughput screening and lead optimization.