一种验证SPR传感器金膜表面固定蛋白质分子有效性的简易途径

Simple Way to Validate the Efficiency of Immobilization of Proteins on Gold Surface in SPR Sensor

利用表面等离子共振传感器进行免疫检测的实验中,需要在化学性质稳定的金膜表面固定蛋白质分子探针。如何验证一种方法是否能够成功固定蛋白质分子具有非常重要的现实意义。提出的验证方法采用酶反应作为标志,如果某种方法确实能有效固定蛋白质分子,那么它也能固定酶。将固定好酶的金膜浸入相应的底物溶液中发生变色反应,测量底物溶液的吸收光谱,即可根据结果判断是否有效固定了酶,同时达到判断这种方法是否可固定蛋白质的目的。具有快速、方便、低成本等优点。采用经过验证的方法固定抗IgG后通入IgG,实验响应良好。

Immobilizing proteins on gold surface is very important for the immunological detection with Surface Plasmon Resonance Sensors. It is practical to investigate how to validate that a method can effectively immobilize proteins on the gold surface. A method using enzyme （IgC-HRP） as a flag is presented. If a method can immobilize proteins, it can also immobilize enzyme. The IgG-HRP linked gold surface is treated with the solution of TMB and H202. The spectrum of the solution can tell whether the enzyme is successfully immobilized and whether that method can effectively immobilize proteins. Terminate the reaction with HCI, there will be a peak at 450nm if the IgG-HRP is indeed immobilized. One immobilizing method was tested in this way. Anti-IgG was immobilized with this tested method, and the result was satisfied after the IgG was combined