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基于成像椭偏仪的生物传感器中蛋白质的共价固定化。

Covalent immobilization of proteins for the biosensor based on imaging ellipsometry.

作者信息

Wang Zhan-Hui, Jin Gang

机构信息

National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080, PR China.

出版信息

J Immunol Methods. 2004 Feb 15;285(2):237-43. doi: 10.1016/j.jim.2003.12.002.

Abstract

In the development of biosensors, the immobilization of biomolecules at interfaces played a crucial role. The feasibility of using 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (Glu) to modify silicon surface to immobilize covalently protein for immunoassay with the biosensor based on imaging ellipsometry was investigated. The higher density and stability of human IgG layer could be obtained on the silicon surface modified with APTES and Glu than that on the silicon surface modified with dichlorodimethylsilane (DDS). The human IgG molecules immobilized covalently on APTES-Glu surface bound more anti-IgG molecules than that on DDS surface, which indicated that the human IgG molecules could maintain higher binding capability on APTES-Glu surface. Tween 20 was able to block the undesirable adsorption on APTES-Glu surface, and also enhanced the recognition between human IgG and its antibody on both APTES-Glu and DDS surfaces. The combination of this protein covalent immobilization and the biosensor has the potential to be developed into a fast, simple immunoassay technique.

摘要

在生物传感器的发展过程中,生物分子在界面的固定化起着至关重要的作用。研究了使用3-氨丙基三乙氧基硅烷(APTES)和戊二醛(Glu)修饰硅表面以共价固定蛋白质,用于基于成像椭偏仪的生物传感器免疫分析的可行性。与用二氯二甲基硅烷(DDS)修饰的硅表面相比,在经APTES和Glu修饰的硅表面上可获得更高密度和稳定性的人IgG层。共价固定在APTES-Glu表面上的人IgG分子比在DDS表面上结合更多的抗IgG分子,这表明人IgG分子在APTES-Glu表面上能够保持更高的结合能力。吐温20能够阻断APTES-Glu表面上不必要的吸附,并且还增强了在APTES-Glu和DDS表面上人IgG与其抗体之间的识别。这种蛋白质共价固定化与生物传感器的结合有潜力发展成为一种快速、简单的免疫分析技术。

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