Zwitterionic polydopamine/protein G coating for antibody immobilization: toward suppression of nonspecific binding in immunoassays

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Title
Zwitterionic polydopamine/protein G coating for antibody immobilization: toward suppression of nonspecific binding in immunoassays
Author(s)
Jihyun Byun; S Cho; Jeong Moon; Hongki Kim; Hyunju Kang; Juyeon JungEun-Kyung LimJinyoung Jeong; H G Park; W K Cho; Taejoon Kang
Bibliographic Citation
Acs Applied Bio Materials, vol. 3, no. 6, pp. 3631-3639
Publication Year
2020
Abstract
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.
Keyword
zwitterionic dopamineprotein Gantibody immobilizationnonspecific bindingimmunoassayinfluenza virussurface-enhanced Raman scattering
ISSN
2576-6422
DOI
http://dx.doi.org/10.1021/acsabm.0c00264
Type
Article
Appears in Collections:
Division of Biomaterials Research > Bionanotechnology Research Center > 1. Journal Articles
Division of Research on National Challenges > Environmental diseases research center > 1. Journal Articles
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