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无阻断且无需基质的血清学微阵列免疫分析

Blocking-Free and Substrate-Independent Serological Microarray Immunoassays.

机构信息

Laboratory for Chemistry and Physics of Interfaces, Department of Microsystems Engineering (IMTEK) , University of Freiburg , Georges-Köhler-Allee 103 , 79110 Freiburg , Germany.

Scienion AG , Volmerstrasse 7b , 12489 Berlin , Germany.

出版信息

Biomacromolecules. 2018 Dec 10;19(12):4641-4649. doi: 10.1021/acs.biomac.8b01334. Epub 2018 Nov 28.

Abstract

In bioanalytical applications, many coating strategies have been established for so-called "blocking" of the surfaces. However, most of the procedures developed so far require additional processing steps for surface blocking and small variations in the blocking efficiency result in increased background noise, which lowers the overall sensitivity of an assay. In this study, we demonstrate the preparation of a bioanalytical surface with a thin film of a photo-cross-linkable copolymer that is transformed photochemically into a surface-attached hydrogel network. The presented coating is directly applicable to various plastic substrates that are used for bioassays without the need for any prior surface modification. Such a strategy allows facile one-step immobilization of biomolecules for bioanalysis and protein-repellent properties for avoiding unspecific adsorption of analyte molecules during the assay. The protein adsorption behavior of the hydrogel-coated and blank surfaces is measured by SPR with human serum and physisorption of labeled detection antibodies. We show that the hydrogel surfaces used lower unspecific background signals and background noise and thus increase the sensitivities of the microarray immunoassays.

摘要

在生物分析应用中,已经建立了许多所谓的“封闭”表面的涂层策略。然而,迄今为止开发的大多数程序都需要额外的表面封闭处理步骤,并且封闭效率的微小变化会导致背景噪声增加,从而降低分析的整体灵敏度。在这项研究中,我们展示了一种具有光交联共聚物的薄膜的生物分析表面的制备,该薄膜可以通过光化学转化为表面附着的水凝胶网络。所提出的涂层可直接应用于各种用于生物分析的塑料基质,而无需任何预先的表面修饰。这种策略允许生物分子的简便一步固定化用于生物分析,并且具有抗蛋白质吸附性,以避免分析过程中分析物分子的非特异性吸附。通过 SPR 用人血清和标记的检测抗体的物理吸附测量了水凝胶涂层和空白表面的蛋白质吸附行为。我们表明,所使用的水凝胶表面降低了非特异性背景信号和背景噪声,从而提高了微阵列免疫分析的灵敏度。

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