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Immunoactive two-dimensional self-assembly of monoclonal antibodies in aqueous solution revealed by atomic force microscopy

Nature Materials volume 13, pages 264–270 (2014)Cite this article

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Abstract

The conformational flexibility of antibodies in solution directly affects their immune function. Namely, the flexible hinge regions of immunoglobulin G (IgG) antibodies are essential in epitope-specific antigen recognition and biological effector function. The antibody structure, which is strongly related to its functions, has been partially revealed by electron microscopy1,2,3,4 and X-ray crystallography5,6, but only under non-physiological conditions. Here we observed monoclonal IgG antibodies in aqueous solution by high-resolution frequency modulation atomic force microscopy7,8 (FM-AFM). We found that monoclonal antibodies self-assemble into hexamers, which form two-dimensional crystals in aqueous solution. Furthermore, by directly observing antibody–antigen interactions using FM-AFM, we revealed that IgG molecules in the crystal retain immunoactivity. As the self-assembled monolayer crystal of antibodies retains immunoactivity at a neutral pH and is functionally stable at a wide range of pH and temperature, the antibody crystal is applicable to new biotechnological platforms for biosensors or bioassays.

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Figure 1: FM-AFM image of a Y-shaped antibody monomer in aqueous solution.
Figure 2: FM-AFM image of flower-like antibody hexamers in aqueous solution.
Figure 3: FM-AFM images of 2D antibody crystals in aqueous solution.
Figure 4: Magnified FM-AFM images of 2D antibody crystals in aqueous solution.
Figure 5: FM-AFM observations of 2D antibody crystals interacting with antigenic or non-antigenic molecules.

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Acknowledgements

This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the SENTAN Program of the Japan Science and Technology Agency, and the Global COE Program of the Japanese Society for the Promotion of Science. S.I. was supported by a Japan Society for the Promotion of Science Fellowship.

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Authors and Affiliations

  1. Department of Electronic Science and Engineering, Kyoto University, Kyoto University Katsura, Nishikyo, Kyoto 615-8510, Japan

    Shinichiro Ido, Kei Kobayashi, Hiroaki Kominami, Kazumi Matsushige & Hirofumi Yamada

  2. Device Solutions Center, Panasonic Corporation, Yagumo-naka-machi 3-1-1, Moriguchi, Osaka 570-8501, Japan

    Hirokazu Kimiya

  3. The Hakubi Center for Advanced Research, Kyoto University, Kyoto University Katsura, Nishikyo, Kyoto 615-8520, Japan

    Kei Kobayashi

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  1. Shinichiro Ido
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Contributions

S.I. performed AFM imaging, analysed data and wrote the paper. H. Kimiya prepared the biological sample and wrote the paper. K.K. developed AFM instruments and electronics and wrote the paper. H. Kominami performed AFM imaging. K.M. designed the study. H.Y. designed the study, developed AFM instruments, analysed the data and wrote the paper. All authors have discussed the results and commented on the manuscript.

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Correspondence to Hirofumi Yamada.

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Ido, S., Kimiya, H., Kobayashi, K. et al. Immunoactive two-dimensional self-assembly of monoclonal antibodies in aqueous solution revealed by atomic force microscopy. Nature Mater 13, 264–270 (2014). https://doi.org/10.1038/nmat3847

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