Oriented and reversible immobilization of His-tagged proteins on two- and three-dimensional surfaces for study of protein-protein interactions by a QCM biosensor
Oriented and reversible immobilization of His-tagged proteins on two- and three-dimensional surfaces for study of protein-protein interactions by a QCM biosensor
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通过 QCM 生物传感器将 His 标记蛋白定向且可逆地固定在二维和三维表面上，用于研究蛋白质-蛋白质相互作用
DOI：
10.1016/j.snb.2015.10.096
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发表时间：
2016-03-01
影响因子：
8.4
通讯作者：
Pei, Zhichao
中科院分区：
文献类型：
--
作者：
Li, Xueming;Song, Siyu;Pei, Zhichao
构建了二维和三维His标签捕获表面，用于在石英晶体微天平（QCM）生物传感器表面上定向可逆固定His标签蛋白质，从而实现对His标签蛋白质与其相互作用蛋白质（分析物）之间相互作用的无标记实时检测。固定在2D Histag捕获表面上的His标记的蛋白质保持比通过胺偶联固定在2D羧基表面上的那些蛋白质更高的结合活性。3D His-标签捕获表面具有2D His-标签捕获表面的约两倍量的固定能力，这使得能够实现更高的检测灵敏度。His标签捕获表面可以任选地再生以去除His标签蛋白以及分析物用于His标签蛋白固定的下一个循环，或者仅选择性地去除分析物，将组氨酸标签蛋白留在表面上用于分析物结合的下一个循环。此外，动力学和亲和性的研究之间的相互作用的His标签的蛋白质和其相互作用的蛋白质进行。本研究提供了一种有效的方法来研究蛋白质-蛋白质相互作用的定向和可逆固定的His标记的蛋白质QCM生物传感器表面。(C)2015 Elsevier B. V.版权所有。
A two-dimensional (2D) and a three-dimensional (3D) His-tag capture surfaces were fabricated for oriented and reversible immobilization of His-tagged proteins on quartz crystal microbalance (QCM) biosensor surfaces, which can be used for label-free and real-time detection of the interactions between His-tagged protein and its interacting protein (analyte). His-tagged proteins immobilized on the 2D Histag capture surface maintained a higher binding activity than those immobilized on a 2D carboxyl surface via amine coupling. The 3D His-tag capture surface has about twice the amount of immobilization capacity as the 2D His-tag capture surface, which enables a higher sensitivity for detection. His-tag capture surface can be optionally regenerated to remove the His-tagged protein as well as the analyte for the next cycle of His-tagged protein immobilization, or to only selectively remove the analyte, leaving the Histagged protein on the surface for the next cycle of analyte binding. Furthermore, the kinetic and affinity studies of the interactions between the His-tagged protein and its interacting protein were performed. This study provides an efficient way to study protein-protein interactions by oriented and reversible immobilization of His-tagged proteins on QCM biosensor surfaces. (C) 2015 Elsevier B.V. All rights reserved.