Immobilization of unraveled immunoglobulin G using well-oriented ZZ-His protein on functionalized microtiter plate for sensitive immunoassay
Immobilization of unraveled immunoglobulin G using well-oriented ZZ-His protein on functionalized microtiter plate for sensitive immunoassay
复制标题
使用定向良好的 ZZ-His 蛋白将未解开的免疫球蛋白 G 固定在功能化微量滴定板上，用于灵敏的免疫测定
DOI：
10.1016/j.ab.2012.09.028
复制
发表时间：
2013-01-15
影响因子：
2.9
通讯作者：
Cheng, Yuan-Zheng
中科院分区：
文献类型：
--
作者：
Yang, Hong-Ming;Liang, Shu-Juan;Cheng, Yuan-Zheng
Highly efficient protein immobilization is extremely crucial for solid-phase immunoassays. We present a strategy for oriented immobilization of functionally intact immunoglobulin G (IgG) on a polystyrene microtiter plate via iminodiacetic acid (IDA)-Ni2+ and ZZ-His protein interaction. We immobilized a ZZ-EAP (Escherichia coli alkaline phosphatase)-His fusion protein, which exhibits Fc binding, His tag, and intrinsic AP activities, and analyzed it against the interaction between rabbit IgG anti-horseradish peroxidase (anti-HRP) and its binding partner HRP to investigate the specificity and efficacy of this method. We compared the IDA-Ni2+-(ZZ-His) method with ZZ-EAP random immobilization using sandwich enzyme-linked immunosorbent assay, and the results showed that the former method had an enhanced signal, 10-fold higher sensitivity, and a wider linear range. Thus, the proposed method allows a broad range of oriented immobilized functionally intact IgG antibodies on polystyrene plates using only one type of IDA-Ni2+ chelate surface because the ZZ protein can bind to the Fc region of various IgGs. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.
