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Zeptomole Detection of an Enzyme by a Simple Colorimetric Method

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Abstract

An enzyme immunoassay, in which an enzyme (e.g., alkaline phosphatase, ALP) is conjugated with an antibody, is a precise and simple protein detection method. Precise measurements of enzymes at low concentrations allow for ultrasensitive protein detection. The application of a phosphorylated substrate to ALP, followed by using a dephosphorylated substrate in thionicotinamide-adenine dinucleotide cycling, provides a simple and precise quantification of ALP. We describe a protocol for detecting ALP at the zeptomole level using a simple colorimetric method.

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Acknowledgments

This work was partly supported by the Waseda University grants for Specific Research Projects (grant numbers 2017A-015 and 2019C-123).

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Authors and Affiliations

  1. Department of Biology, Waseda University, 2-2 Wakamatsucho Shinjuku, Tokyo, 162-8480, Japan

    Kanako Iha, Yuta Kyosei, Mayuri Namba, Daiki Makioka, Sou Yamura & Etsuro Ito

  2. Waseda Research Institute for Science and Engineering, Waseda University, 3-4-1 Okubo Shinjuku, Tokyo, 169-8555, Japan

    Satoshi Watabe & Etsuro Ito

  3. School of Pharmaceutical Sciences, Health Sciences University of Hokkaido, 1757 Kanazawa Tobetsu, Ishikari, Hokkaido, 061-0293, Japan

    Teruki Yoshimura

  4. Graduate Institute of Medicine, Kaohsiung Medical University, No. 100 Shih-Chuan 1st Rd. Sanmin, Kaohsiung, 80708, Taiwan

    Etsuro Ito

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  1. Kanako Iha
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  2. Yuta Kyosei
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  3. Mayuri Namba
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  4. Daiki Makioka
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  5. Sou Yamura
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  6. Satoshi Watabe
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  7. Teruki Yoshimura
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  8. Etsuro Ito
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Correspondence to Etsuro Ito.

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Iha, K., Kyosei, Y., Namba, M. et al. Zeptomole Detection of an Enzyme by a Simple Colorimetric Method. ANAL. SCI. 37, 1469–1472 (2021). https://doi.org/10.2116/analsci.21N009

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  • DOI: https://doi.org/10.2116/analsci.21N009

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