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High-density immobilization of antibodies onto nanobead-coated cyclic olefin copolymer plastic surfaces for application as a sensitive immunoassay chip

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Abstract

Our research efforts have been devoted to development of nanobead multilayer-based sensitive immunoassays on cyclic olefin copolymer (COC) plastic surfaces. To facilitate nanobead attachment and impart antibiofouling properties to a COC substrate, we used an amphiphilic copolymer comprising benzyl, polyethylene glycol, and reactive ester moieties to coat the hydrophobic COC surface in an aqueous environment. Subsequently, NH2-modified polystyrene nanobeads were reacted with the polymer-coated COC surface and further assembled into multilayers that increased the overall surface area available for attaching capture antibodies. After treatment of the nanobead multilayers with an amine-reactive homobifunctional crosslinker, a model capture antibody (anti-rabbit IgG) was covalently immobilized onto the activated surface of nanobeads. Finally, a sandwich immunoassay was carried out using rabbit IgG as a target analyte and rhodamine-labeled anti-rabbit IgG as a probe. Compared with a nanobead-free, polymer-coated COC surface, the nanobead multilayer-based immunoassay exhibited ~4-fold higher fluorescence intensity. In addition, our nanobead-based assay system exhibited a wide dynamic range of detection (0.1 to 1,000 ng/mL) and high specificity for rabbit IgG. Furthermore, much better detection sensitivity for rabbit IgG was attained in the nanobead multilayer-based immunoassay than with a conventional ELISA system (0.1 ng/mL versus 10 ng/mL), indicating the potential value of the proposed immunoassay system in plastic-based portable biochip applications.

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Acknowledgements

This study was supported by a grant from the National R&D Program for Cancer Control, Ministry for Health, Welfare, and Family Affairs, Republic of Korea and by WCU (World Class University) program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (grant number: R31-10071).

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Authors and Affiliations

  1. Department of Medical System Engineering, Institute of Medical System Engineering, Gwangju Institute of Science and Technology (GIST), 123 Cheomdan-gwagiro, Buk-gu, Gwangju, 500-712, Republic of Korea

    Daekyung Sung

  2. Department of Medical System Engineering, Gwangju Institute of Science and Technology (GIST), 123 Cheomdan-gwagiro, Buk-gu, Gwangju, 500-712, Republic of Korea

    Sung Yang

  3. Department of Nanobio Materials and Electronics, Gwangju Institute of Science and Technology (GIST), 123 Cheomdan-gwagiro, Buk-gu, Gwangju, 500-712, Republic of Korea

    Sung Yang

  4. School of Mechatronics, Gwangju Institute of Science and Technology (GIST), 123 Cheomdan-gwagiro, Gwangju, 500-712, Republic of Korea

    Sung Yang

  5. BioHealth IT Convergence Research Department, IT Convergence Technology Research Laboratory, Electronics and Telecommunications Research Institute (ETRI), 218 Gajeong-ro, Yuseong-gu, Daejeon, 305-700, Republic of Korea

    Jeong Won Park

  6. KAIST Institute for the Biocentury, Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Daejeon, 305-701, Republic of Korea

    Sangyong Jon

  7. Graduate School of Nanoscience and Technology (WCU), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Daejeon, 305-701, Republic of Korea

    Sangyong Jon

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  1. Daekyung Sung
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  2. Sung Yang
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  3. Jeong Won Park
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Correspondence to Sangyong Jon.

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Sung, D., Yang, S., Park, J.W. et al. High-density immobilization of antibodies onto nanobead-coated cyclic olefin copolymer plastic surfaces for application as a sensitive immunoassay chip. Biomed Microdevices 15, 691–698 (2013). https://doi.org/10.1007/s10544-012-9732-x

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  • DOI: https://doi.org/10.1007/s10544-012-9732-x

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