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High-performance affinity beads for identifying drug receptors

Nature Biotechnology volume 18, pages 877–881 (2000)Cite this article

Abstract

We have developed a method using novel latex beads for rapid identification of drug receptors using affinity purification. Composed of a glycidylmethacrylate (GMA) and styrene copolymer core with a GMA polymer surface, the beads minimize nonspecific protein binding and maximize purification efficiency. We demonstrated their performance by efficiently purifying FK506-binding protein using FK506-conjugated beads, and found that the amount of material needed was significantly reduced compared with previous methods. Using the latex beads, we identified a redox-related factor, Ref-1, as a target protein of an anti-NF-κB drug, E3330, demonstrating the existence of a new class of receptors of anti-NF-κB drugs. Our results suggest that the latex beads could provide a tool for the identification and analysis of drug receptors and should therefore be useful in drug development.

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Figure 1: Purification of FKBP.
Figure 2: Purification of the E3330 receptors.
Figure 3: Identification of the E3330-binding protein as Ref-1.
Figure 4: In vitro binding assay using SPR.
Figure 5: Functional interaction of Ref-1 with the NF-κB p50 subunit.
Figure 6: Effects of Ref-1 and E3330 on NF-κB-dependent gene expression.

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Acknowledgements

The authors are grateful to Eisai Co., Ltd. for providing E3330 and NH2-E3330. This work was supported in part by a research grant from Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Cooperation (J.S.T.), Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science, Sports and Culture, and a grant of R and D Projects in Cooperation with Academic Institutions from New Energy and Industrial Technology Development Organization (NEDO). We thank Dr. David Buchanan for critical reading of the manuscript.

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Authors and Affiliations

  1. Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, 226-8501, Japan

    Noriaki Shimizu, Kotaro Sugimoto, Jianwei Tang, Takeyuki Nishi, Iwao Sato, Tatsufumi Yoshikawa, Fumihiko Suzuki, Hajime Watanabe & Hiroshi Handa

  2. Department of Anatomy, Nihon University School of Medicine, Tokyo, 173-8601, Japan

    Masaki Hiramoto & Shin Aizawa

  3. Frontier Collaborative Research Center, Tokyo Institute of Technology, Yokohama, 226-8503, Japan

    Mamoru Hatakeyama, Reiko Ohba, Hideaki Hatori & Hiroshi Handa

  4. Institute of Life Sciences, Mitsubishi Chemical Corporation, Machida, 194-8511, Japan

    Akira Oomori

  5. Institute of Medical Science, University of Tokyo, Tokyo, 108-8639, Japan

    Hirotoshi Tanaka

  6. Faculty of Science and Technology, Keio University, Yokohama, 223-8522, Japan

    Haruma Kawaguchi

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  1. Noriaki Shimizu
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Correspondence to Hiroshi Handa.

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Shimizu, N., Sugimoto, K., Tang, J. et al. High-performance affinity beads for identifying drug receptors. Nat Biotechnol 18, 877–881 (2000). https://doi.org/10.1038/78496

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