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An Open Sandwich Immunochromatography for Non-competitive Detection of Small Antigens

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Abstract

Immunochromatography assay is an easy and rapid on-site detection method. However, conventional sandwich immunochromatographies using two antibodies can only detect target molecules above a threshold size. Small molecules below 1000 in molecular weight are usually detected using competitive immunoassay. However, competitive immunoassay is not suitable for visual detection of low concentration samples. Based on the principles of open sandwich immunoassay, which detects small molecules via interchain interaction of separated variable region fragments (VH and VL) from a single antibody, we developed non-competitive open sandwich immunochromatography. Bone Gla protein (BGP)-C7, a peptide containing the seven C-terminal amino acids of human osteocalcin, was selected as the target. By using VL fragments fixed on a nitrocellulose membrane, and colored cellulose bead-labeled VH fragments, we specifically detected 10 ng/mL of BGP-C7. This is the first report of open sandwich immunochromatography, which is an easy and rapid method for on-site, signal-on detection of small molecules.

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Acknowledgments

We thank Dr. Tetsuya Kitaguchi for his valuable suggestions. This work was supported in part by the JSPS KAKENHI Grant Number JP18H03851 from the Japan Society for the Promotion of Science, and by JST-Mirai Grant Number JPMJMI18D9 from the Japan Science and Technology Agency. We thank the Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials from MEXT, Japan, for the support provided for our studies.

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Authors and Affiliations

  1. Food Research Center, Central Laboratory, Nippon Flour Mills Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa, 243-0031, Japan

    Yoshitaka Harada & Mika Tsuna

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

    Yoshitaka Harada, Yuki Ohmuro-Matsuyama & Hiroshi Ueda

Authors
  1. Yoshitaka Harada
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  2. Yuki Ohmuro-Matsuyama
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  3. Mika Tsuna
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  4. Hiroshi Ueda
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Correspondence to Hiroshi Ueda.

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Harada, Y., Ohmuro-Matsuyama, Y., Tsuna, M. et al. An Open Sandwich Immunochromatography for Non-competitive Detection of Small Antigens. ANAL. SCI. 37, 455–459 (2021). https://doi.org/10.2116/analsci.20SCP06

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  • DOI: https://doi.org/10.2116/analsci.20SCP06

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