A highly sensitive assay for protein with dibromochloro-arsenazo-Al3+ using resonance light scattering technique and its application
A highly sensitive assay for protein with dibromochloro-arsenazo-Al3+ using resonance light scattering technique and its application
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DOI：
10.1016/s0039-9140(02)00075-9
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发表时间：
2002-06-10
期刊：
影响因子：
6.1
通讯作者：
Hu, ZD
中科院分区：
文献类型：
--
作者：
Jia, RP;Dong, LJ;Hu, ZD
建立了一种简单、灵敏、选择性好的共振光散射（RLS）法测定蛋白质的方法。该方法基于蛋白质与偶氮胂-DBC-Al ~（3+）在pH 5.0-7.0范围内的相互作用，在300-550 nm波长范围内，偶氮胂-DBC-Al ~（3+）的共振散射信号显著增强，共振光散射最大平台在405-420 nm。该方法可测定2.50-50.00 μ g/ml牛血清白蛋白（BSA）和2.50-60.00 μ g/ml人血清白蛋白（HSA），检出限为标准偏差的3倍。在6次空白测定中，BSA和HSA的平均浓度分别为123.4和89.6 ng ml（-1）。此外，该方法不受许多氨基酸和金属离子的干扰。该方法具有灵敏度高、选择性好、重现性好等优点，已成功应用于人血清样品中总蛋白的测定。机理研究表明，偶氮胂-DBC-Al ~（3+）与BSA的结合主要依赖于静电力，从而增强了偶氮胂-DBC-Al ~（3+）-蛋白体系的共振光散射。(C)2002 Elsevier Science B. V.保留所有权利。
A simple, sensitive and selective method has been developed for the determination of protein using resonance light scattering (RLS) technique. The method is based on the interaction of protein and arsenazo-DBC-Al3+ in the pH range of 5.0-7.0, which causes a substantial enhancement of the resonance scattering signal of arsenazo-DBC-Al3+ in the wavelength range of 300-550 nm with the maximum RLS platform at 405-420 nm. With this method, 2.50-50.00 mug ml(-1) of bovine serum albumin (BSA) and 2.50-60.00 mug ml(-1) of human serum albumin (HSA) can be determined, and the detection limits, calculated three times the standard deviation (S.D.) of six blank measurements, for BSA and HSA were 123.4 and 89.6 ng ml(-1), respectively. Moreover, the method is free from interference from many amino acids and metal ions. The method, with high sensitivity, selectivity and reproducibility, was satisfactorily applied to the determination of total protein in human serum samples. Mechanism studies indicated that arsenazo-DBC-Al3+ could bind to BSA depending mainly on electrostatic forces, which results in enhanced RLS in the arsenazo-DBC-Al3+-protein system. (C) 2002 Elsevier Science B.V. All rights reserved.