Blocking-Free and Substrate-Independent Serological Microarray Immunoassays,Biomacromolecules

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Blocking-Free and Substrate-Independent Serological Microarray Immunoassays
Biomacromolecules ( IF 5.5 ) Pub Date : 2018-11-15 00:00:00 , DOI: 10.1021/acs.biomac.8b01334
Frank D. Scherag ₁ , Andreas Mader ₂ , Marc Zinggeler ₁ , Nicole Birsner ₁ , Richard E. Kneusel ₃ , Thomas Brandstetter ₁ , Jürgen Rühe ₁

Affiliation

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.

中文翻译：

无阻塞和独立于底物的血清微阵列免疫测定

在生物分析应用中，已经建立了许多用于所谓的“阻塞”表面的涂层策略。但是，迄今为止开发的大多数程序都需要用于表面封闭的附加处理步骤，并且封闭效率的微小变化会导致背景噪声增加，从而降低了测定的总体灵敏度。在这项研究中，我们证明了具有光可交联共聚物薄膜的生物分析表面的制备，该薄膜可通过光化学方式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用于生物测定的各种塑料基材，而无需任何先前的表面修饰。这样的策略允许生物分子的容易的一步固定化以用于生物分析和蛋白排斥性质，从而避免在测定期间非特异性吸附分析物分子。用人血清通过SPR和标记的检测抗体的物理吸附来测量水凝胶涂层和空白表面的蛋白质吸附行为。我们表明水凝胶表面使用较低的非特异性背景信号和背景噪声，从而增加了微阵列免疫测定的灵敏度。

更新日期：2018-11-15

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