1)深圳大学物理与光电工程学院,光电器件与系统教育部/广东省重点实验室,广东深圳 518060; 2)湖南省计量检测研究院,湖南长沙 410014

光学; 表面等离子共振传感; 动力学分析; 蛋白A; 蛋白G; 抗体

The kinetic study on the interactions of IgG and antibody-binding proteins based on SPR sensor
CAI Zhiwen1, XIAO Xiaoping2, WANG Xueliang1, SHAO Yonghong1, and ZHOU Jie1

1)College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China2)Hunan Institute of Metrology and Test, Changsha 410014, Hunan Province, P.R.China

optics; surface plasmon resonance sensing; kinetic analysis; protein A; protein G; antibody

DOI: 10.3724/SP.J.1249.2021.01098


将自研的基于波长扫描的表面等离子体共振传感系统与动力学分析方法相结合,研究葡萄球菌蛋白A、链球菌蛋白G与免疫球蛋白G(immunoglobulin G, IgG)的相互作用关系.实验测得,蛋白A和蛋白G与IgG的结合速率常数分别为1.3×105 L·mol-1·s-1和5.0×104 L·mol-1·s-1,说明蛋白A与IgG的结合效率更高.同时测得蛋白A和蛋白G与IgG的解离速率常数分别为2.1×10-2 s-1和5.0×10-3 s-1,说明蛋白G与IgG形成的复合物更稳定.此外,由解离速率常数与结合速率常数的比值得出蛋白G对IgG的亲和力较大.实验结果可为优化抗体固定方法提供必要参考和理论支撑.

We study the interactions between staphylococal protein A or streptococcus protein G and immunoglobulin IgG, respectively, based on the self-developed wavelength-based surface plasmon resonance(SPR)sensing system combined with the kinetic analysis. The experimental results show that the association rate constants ka of protein A and protein G with IgG are 1.3×105 L·mol-1·s-1 and 5.0×104 L·mol-1·s-1, indicating that the protein A binds to IgG more efficiently. At the same time, the dissociation rate constants kd of protein A and protein G from IgG are detected to be 2.1×10-2 s-1 and 5.0×10-3 s-1, indicating that the complex formed by protein G and IgG is more stable. In addition, the ratio of kd to ka indicates that protein G has higher affinity for IgG. This study may provide necessary reference and important theoretical support for the optimization of antibody immobilization methods.