用共振光散射技术研究蛋白质与丽春红2R的相互作用

Study on the Interaction of Proteins with Ponceau 2R by the Resonance Rayleigh Light-Scattering Technique and Its Analytical Applications

在pH3.50的Britton Robinson缓冲溶液中,丽春红2R与蛋白质发生结合反应,使最大波长为352.5nm的共振散射光谱得到加强。据此建立了利用丽春红2R作探针,用共振散射光谱技术测定蛋白质的新方法。散射光强度与牛血清白蛋白(BSA)、人血清白蛋白(HAS)和免疫球蛋白(IgG)的浓度在0.25～17.5μg/mL范围内成正比。方法的稳定性好,快速、简便,绝大多数氨基酸、金属离子均不产生干扰,用于人血清样品中蛋白质的测定,获得了满意的结果,加标回收率为93.4%～100.1%。

In Britton-Robinson buffer of pH 3.50, the binding reaction of ponceau 2R with proteins enhances remarkably the resonance Rayleigh light-scattering signal at maximum wavelength of 352.5 nm. Based on this phenomenon, a new method for the determination of proteins by the resonance Rayleigh light-scattering technique(RRLS) using ponceau 2R as probe agent has been developed. The light-scattering intersity is proportional to the concentration of bovine serum albumin (BSA), human serum albumin(HAS) and human Immunoglobulin G(IgG) in the range 0.25～17.5 μg/mL. The method is stable, rapid and simple, and free of the interference from most of amino acids, metal ions and other foreign substances. It has been used to determine total proteins in human serum samples with satisfactory result. The recoveries are in the range 93.4%～100.1%.