蛋白质与铕（III）-金霉素络合物之间的相互作用研究及荧光分析法测定蛋白质

Study on the interaction between protein and Eu(III)-chlorotetracycline complex and the determination of protein using the fluorimetric method.

作者信息

Liu Shufang, Yang Jinghe, Wu Xia, Wang Fei, Wang Feng, Jia Zhen, Mao Lijuan

机构信息

Key Laboratory of Colloid and Interface Chemistry (Shandong University), Ministry of Education, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, People's Republic of China.

出版信息

Luminescence. 2004 Nov-Dec;19(6):352-7. doi: 10.1002/bio.792.

DOI:10.1002/bio.792

PMID:15558687

Abstract

Chlorotetracycline (CTC) can react with europium ions Eu3+, and the complex emits the intrinsic fluorescence of Eu3+. The intensity is greatly enhanced by proteins and this forms the basis of a new fluorimetric method for determination of protein. Further research indicates that under optimum conditions, the enhanced intensity of fluorescence is in proportion to the concentration of proteins, in the range 2.0 x 10(-7)-1.0 x 10(-5) g/mL for bovine serum albumin (BSA) (linear equation, I(f) = 34.35933 + 11.54467 x 10(6)C)(r = 0.99895) and 8.0 x 10(-7)-1.0 x 10(-5) g/mL for human serum albumin (HSA) (linear equation, I(f) = 76.58881 + 5.3569 x 10(6)C) (r = 0.99283). Detection limits (S/N = 3) were 8.9 x 10(-9) g/mL for BSA and 3.3 x 10(-8) g/mL for HSA. In an assay for BSA in calf serum, this method gave a value close to that determined by the UV spectrophotometric method.

摘要

金霉素（CTC）能与铕离子Eu3+发生反应，该络合物会发出Eu3+的固有荧光。蛋白质可使荧光强度大幅增强，这构成了一种测定蛋白质的新型荧光分析法的基础。进一步研究表明，在最佳条件下，荧光增强强度与蛋白质浓度成正比，对于牛血清白蛋白（BSA），浓度范围为2.0×10(-7)-1.0×10(-5) g/mL（线性方程，I(f)=34.35933 + 11.54467×10(6)C）（r = 0.99895）；对于人血清白蛋白（HSA），浓度范围为8.0×10(-7)-1.0×10(-5) g/mL（线性方程，I(f)=76.58881 + 5.3569×10(6)C）（r = 0.99283）。牛血清白蛋白的检测限（S/N = 3）为8.9×10(-9) g/mL，人血清白蛋白的检测限为3.3×10(-8) g/mL。在小牛血清中牛血清白蛋白的测定中，该方法得到的值与紫外分光光度法测定的值相近。