实时生物分子相互作用分析技术用于链霉亲和素-生物素化抗体的层层组装研究

Layer-by-Layer Assembly of Multilayer Films Composed of Streptavidin and Biotinylated Antibody by Real-time Biomolecular Interaction Analysis

使用生物分子相互作用分析 ( Biomolecular interaction analysis,BIA)技术实时监测了在链霉亲和素表面层层组装亲和素 -生物素化抗体多层膜的过程 ,结果表明 ,通过链霉亲和素与生物素之间的强亲和作用 ,能够在表面形成均一的多层膜 ,并用实时 BIA技术求得了每层蛋白质的表面浓度 .对于生物素化抗体 ,单层吸附表面浓度为 1 .32 ng/mm2 ;对于链霉亲和素 ,单层吸附表面浓度为 2 .93ng/mm2 .

Layer by layer assembly of multilayer films of streptavidin and biotinylated antibody was completed on the streptavidin coated surface. Real time biomolecular interaction analysis(BIA) based on surface plasmon resonance technique was used to monitor the multilayer assembly in solution continuously. The results indicate that the uniform multilayer film can be fabricated successfully based on the strong interaction between streptavidin and biotin. The mean surface mass concentration of each adsorption layer is 1 32 ng/mm 2 for biotinylated antibody, 2 93 ng/mm 2 for streptavidin, according to the correlation of SPR response with surface concentration.