利用表面等离子体谐振生物传感器实时监测酶促反应的新方法

A Novel Method for Detecting and Analyzing the Enzyme Catalytic Reactions with Surface Plasmon Resonance Biosensor

在理论上 ,基于表面等离子体谐振 (surfaceplasmonresonance,SPR)生物传感器的酶促反应检测分析方法被证明是可行的。这种方法与通常SPR传感器检测技术不同 ,它消除通常当成检测目标的表面吸附信号 ,而检测通常认为是噪声的本体折射率变化信号 ,以此实时检测酶促反应的动态过程。利用自产的SPR 2 0 0 0生化分析仪 ,在检测胶原酶Ⅰ降解胶原Ⅰ的实验中检测到了反应引起的反应液本体折射率的上升变化 ,从而证明了这种新颖的检测方法实际上也是可行的。这种技术一旦成熟 ,可以用来进行酶学分析及酶靶标的药物筛选研究开发。

Based on surface plasmon resonance (SPR) biosensor, a novel detection and kinetic analysis method for analyzing enzyme catalytic reactions was proved to be feasible in theory. In contrast to traditional SPR detection and analysis method, this method monitoring an enzyme catalytic reaction by removing surface adsorption signals and detecting the SPR signals caused by the bulk reflective index changes in the enzyme-substrate solution. With a home-made SPR equipment SPR-2000, and detecting the break-down reaction between collagenase type I and collagen type I by this method, a rising SPR curve was observed which revealed the increase of the solution bulk refractive index caused by the reaction. Therefore, this method is also feasible in practice. The mature form of this method would be powerful in enzyme assay and drug screening based on enzyme targets.