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Open sandwich ELISA: A novel immunoassay based on the interchain interaction of antibody variable region

Nature Biotechnology volume 14, pages 1714–1718 (1996)Cite this article

Abstract

We describe an immunoassay that is based on the interchain interaction of separated VL and VH chains from a single chain antibody variable region. In the presence of antigen, the chains reassociate. VL fragments of anti-hen egg lysozyme (HEL) antibody HyHEL-10 were immobilized on microtiter plates. Samples were coincubated with an M13-displayed VH chain, and assayed with peroxidase-labeled anti-M13 antibody. Signal was detected in direct proportion to the amount of HEL in the sample. Wide dynamic range with <15 ng/ml sensitivity was attained.

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Author notes

  1. Hiroshi Ueda: email: hueda@bio.t.u-tokyo.ac.jp

Authors and Affiliations

  1. Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113, Japan

    Hiroshi Ueda, Kazuishi Kubota, Eiji Suzuki, Teruyuki Nagamune & Hajime Nishimura

  2. Department of Biochemistry and Engineering, Faculty of Engineering, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-77, Japan

    Kouhei Tsumoto & Izumi Kumagai

  3. Boeringer Mannheim Corporation, 2400 Bisso Lane, Concord, CA, 94520-4117, USA

    Paula A. Schueler & Walter C. Mahoney

  4. Medical Research Council Centre for Protein Engineering, Hills Rd., Cambridge, CB2 2QH, UK

    Greg Winter

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Ueda, H., Tsumoto, K., Kubota, K. et al. Open sandwich ELISA: A novel immunoassay based on the interchain interaction of antibody variable region. Nat Biotechnol 14, 1714–1718 (1996). https://doi.org/10.1038/nbt1296-1714

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  • DOI: https://doi.org/10.1038/nbt1296-1714

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