Wang Zhanhui, Jin Gang
National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, 15, Bei-Si-Huan West Road, Beijing 100080, PR China.
J Biochem Biophys Methods. 2003 Sep 30;57(3):203-11. doi: 10.1016/s0165-022x(03)00109-x.
The feasibility of using protein A to immobilize antibody on silicon surface for a biosensor with imaging ellipsometry was presented in this study. The amount of human IgG bound with anti-IgG immobilized by the protein A on silicon surface was much more than that bound with anti-IgG immobilized by physical adsorption. The result indicated that the protein A could be used to immobilize antibody molecules in a highly oriented manner and maintain antibody molecular functional configuration on the silicon surface. High reproducibility of the amount of antibody immobilization and homogenous antibody adsorption layer on surfaces could be obtained by this immobilization method. Imaging ellipsometry has been proven to be a fast and reliable detection method and sensitive enough to detect small changes in a molecular monolayer level. The combination of imaging ellipsometry and surface modification with protein A has the potential to be further developed into an efficient immunoassay protein chip.
本研究展示了利用蛋白A在硅表面固定抗体以构建用于成像椭偏仪的生物传感器的可行性。通过蛋白A固定在硅表面的抗IgG所结合的人IgG量,远多于通过物理吸附固定的抗IgG所结合的人IgG量。结果表明,蛋白A可用于以高度定向的方式固定抗体分子,并在硅表面维持抗体分子的功能构型。通过这种固定方法,可实现抗体固定量的高重现性以及表面均匀的抗体吸附层。成像椭偏仪已被证明是一种快速可靠的检测方法,灵敏度足以检测分子单层水平的微小变化。成像椭偏仪与蛋白A表面修饰的结合有潜力进一步发展成为一种高效的免疫分析蛋白芯片。