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Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains

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Synthetic Protein Switches

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1596))

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Abstract

Antibody-based molecular switches that are able to recognize a range of exogenous antigens can be highly useful as a versatile biosensor. However, regulating the catalytic activity of enzymes by antibodies is still hard to achieve. Here, we describe a design method of unique antibody variable region Fv introduced with two circular permutations, called Clampbody. By tethering the Clampbody to a circularly permuted TEM-1 β-lactamase (BLA), we successfully constructed a genetically encoded molecular switch Cbody-cpBLA that shows antigen-dependent catalytic activity.

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Acknowledgment

We thank Tatsuya Shinoda of Kyowa Medex Co. for generously allowing the use of the genes derived from KTM-219 IgG. This study was supported by Grant-in-Aid for Scientific Research (Grants 24360336 and 15H04191 to H.U. and Grant 46420793 to J.D.) from JSPS, Japan, and partly by Strategic International Collaborative Research Program, Japan Science and Technology Agency (JST).

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Authors and Affiliations

  1. Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8656, Japan

    Hiroto Iwai & Miki Kojima-Misaizu

  2. Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, 4259-R1-18 Nagatsuta-cho, Midori-ku, Yokohama, 226-8503, Japan

    Jinhua Dong & Hiroshi Ueda

  3. College of Chemistry and Chemical Engineering, Linyi University, Shandong, 27600, China

    Jinhua Dong

Authors
  1. Hiroto Iwai
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  2. Miki Kojima-Misaizu
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  3. Jinhua Dong
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  4. Hiroshi Ueda
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Corresponding author

Correspondence to Hiroshi Ueda .

Editor information

Editors and Affiliations

  1. Fachbereich Biologie, Technische Universität Darmstadt, Darmstadt, Germany

    Viktor Stein

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Iwai, H., Kojima-Misaizu, M., Dong, J., Ueda, H. (2017). Creation of Antigen-Dependent β-Lactamase Fusion Protein Tethered by Circularly Permuted Antibody Variable Domains. In: Stein, V. (eds) Synthetic Protein Switches. Methods in Molecular Biology, vol 1596. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6940-1_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6940-1_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6938-8

  • Online ISBN: 978-1-4939-6940-1

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