Zwitterionic Polydopamine/Protein G Coating for Antibody Immobilization: Toward Suppression of Nonspecific Binding in Immunoassays,ACS Applied Bio Materials

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Zwitterionic Polydopamine/Protein G Coating for Antibody Immobilization: Toward Suppression of Nonspecific Binding in Immunoassays
ACS Applied Bio Materials ( IF 4.6 ) Pub Date : 2020-04-20 , DOI: 10.1021/acsabm.0c00264
Jihyun Byun ₁ , Soojeong Cho ₂ , Jeong Moon _{1,

3} , Hongki Kim ₁ , Hyunju Kang ₁ , Juyeon Jung _{1,

4} , Eun-Kyung Lim _{1,

4} , Jinyoung Jeong _{4,

5} , Hyun Gyu Park ₃ , Woo Kyung Cho ₂ , Taejoon Kang ₁

Affiliation

For the development of immunoassays into sophisticated analyte-sensing methods, it is a priority to suppress nonspecific binding in immunoassays. Herein, we report a one-step surface coating method that can not only optimally immobilize antibodies but also suppress nonspecific binding. Zwitterionic dopamine (ZW-DOPA) exhibits distinct antifouling performance, and protein G enables an antibody to have an optimal orientation. A mixture of ZW-DOPA and protein G can be simply coated onto various kinds of surfaces, and the antibody can be immobilized onto the ZW-DOPA/protein G-coated surfaces. The antifouling property of the zwitterionic group, surface-independent coating property of the catechol and amine groups, and antibody-retaining property of protein G synergistically contribute to surface-independent and oriented immobilization of antibodies without nonspecific binding. The surface characteristics of ZW-DOPA/protein G-coated substrates were analyzed by X-ray photoelectron spectroscopy, contact angle goniometry, atomic force microscopy, and ellipsometry. Importantly, the ZW-DOPA/protein G-coated substrates showed high resistance to nonspecific protein adhesion. We also verified that antibodies could be immobilized onto ZW-DOPA/protein G-coated substrates using fluorescence and biolayer interferometry systems. Finally, ZW-DOPA/protein G-coated substrates were employed as immune substrates for influenza virus detection via the naked eye and surface-enhanced Raman scattering, allowing us to efficiently identify the virus. It is anticipated that the developed ZW-DOPA/protein G coating method will be useful for the advancement of immunoassays.

中文翻译：

用于抗体固定的两性离子聚多巴胺/蛋白 G 涂层：抑制免疫测定中的非特异性结合

为了将免疫测定发展为复杂的分析物传感方法，抑制免疫测定中的非特异性结合是当务之急。在这里，我们报告了一种一步表面涂层方法，该方法不仅可以最佳地固定抗体，还可以抑制非特异性结合。两性离子多巴胺 ( ZW -DOPA) 具有独特的防污性能，蛋白质 G 使抗体具有最佳方向。ZW -DOPA 和蛋白 G的混合物可以简单地包被到各种表面上，抗体可以固定在ZW上-DOPA/蛋白 G 涂层表面。两性离子基团的防污性能、邻苯二酚和胺基团的表面独立涂层特性以及蛋白 G 的抗体保留特性协同有助于抗体的表面独立和定向固定，而没有非特异性结合。通过 X 射线光电子能谱、接触角测角法、原子力显微镜和椭偏仪分析ZW -DOPA/蛋白 G 涂层基材的表面特性。重要的是，ZW -DOPA/蛋白 G 包被的底物表现出对非特异性蛋白粘附的高抗性。我们还验证了抗体可以固定在ZW上-使用荧光和生物层干涉测量系统的多巴/蛋白 G 涂层基材。最后，ZW -DOPA/蛋白 G 包被的底物被用作通过肉眼和表面增强拉曼散射检测流感病毒的免疫底物，使我们能够有效地识别病毒。预计开发的ZW -DOPA/蛋白 G 涂层方法将有助于免疫测定的进步。

更新日期：2020-04-20

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