用铀试剂Ⅰ共振光散射法测定蛋白质的研究

Study on Determination of Protein with ArsenazoⅠ Using Resonance Light Scattering Technique

本文提出一种新的光散射探针染料———铀试剂Ⅰ (ArsenazoⅠ ) ,在pH 3 2 9的条件下 ,ArsenazoⅠ本身只有极弱的光散射 ,它与蛋白质的结合物有强烈的共振光散射作用。在λ =4 0 0nm处 ,光散射有最大强度 ,光散射强度与蛋白质的浓度成正比。据此建立了一种测定蛋白质的新方法。该方法简便、快速、灵敏度高 ,对HSA的检测限达到 6 0ng·mL-1 ,线性范围达到 0～ 18μg·mL-1 ,用于人体血清样品的分析并同考马斯亮蓝法比较 ,取得了令人满意的结果。本文也研究了牛血清白蛋白 (BSA)、γ球蛋白、鸡蛋白蛋白、溶菌酶、胃蛋白酶与染料ArsonazoⅠ之间的作用 。

A new RLS (Resonance light scattering) probe is presented in this paper In the Britton Robinson buffer at pH 3 29, ArsenazoⅠ combines with proteins by intermolecular forces, resulted in an enhanced Rayleigh light scattering The RLS intensity reaches maximum at 400 nm and is proportional to the concentration of protein A novel method for the determination of micro amount of protein is developed The assay is simple, rapid and sensitive with a detection limit at 60 ng·mL -1 and a linear range up to 18 μg·mL -1 This method is applied to the determination of human serum protein, and the results, compared to Bradford method, is satisfactory The interaction of ArsenazoⅠ with BSA, γ G, oval albumin, lysozyme, pepsin are investigated and the mechanism of RLS is also discussedthedeterminationofmicro amountofproteinisdev