四羧基镍酞菁共振散射法测定蛋白质

Determination of Protein with Tetra-carboxylic Nickel Phthalocyanine by Resonance Light Scattering

中文关键词:  共振散射,四羧基镍酞菁,蛋白质

英文关键词:resonance light scattering,tetra-carboxylic nickel phathlocyanine,protein

中文摘要:

      建立了一种测定蛋白质的新方法.在pH 3.6的Britton-Robinson(B-R)缓冲溶液中，蛋白质与四羧基镍酞菁NiPc(COOH)₄发生相互作用，使体系在λ=388 nm处的共振散射(RLS)增强，并且增强的散射强度(IRLS)与蛋白质的含量成比例, 据此利用四羧基镍酞菁NiPc(COOH)₄为光谱探针共振散射法测定人血清中的总蛋白质，同时优化了体系光散射检测的实验参数.在最佳的实验条件下，对牛血清白蛋白(BSA)、人血清白蛋白(HSA)、人血清总蛋白(TP)的线性范围分别为0.00～1.20、0.00～1.00、0.00～1.00 mg/L，相应检测限分别为5.97×10^-4 mg/L、2.90×10^-4 mg/L、4.76×10^-4 mg/L.将该方法应用于实际人血清样品中总蛋白的测定，结果与考马斯亮蓝法比较，令人满意.

英文摘要:

      A new method for determination of protein was developed. At pH 3.6 Britton\|Robision buffer solution, the interaction between tetra-carboxylic nickel phathlocyanine and proteins yielded strongly enhanced RLS signals at λ=388 nm and the enhanced intensity of RLS was proportional to the concentration of proteins. Based on this, tetra-carboxylic nickel phathlocyanine was used as a probe to determine the total proteins in human serum samples. Under optimal conditions, the linear range is 0.00～1.20 mg/L for BSA、0.00～1.00 mg/L for HSA、0.00～1.00 mg/L for total protein , respectively, with detection limit of 5.97×10^-4 mg/L for BSA、2.90×10^-4 mg/L for HSA、4.76×10^-4 mg/L for total proteins .The method has been applied to determine the total proteins in human serum samples and the results was compared to the coomassie brilliant blue G-250 method with satisfactory results.