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Noncompetitive Immunodetection of Benzaldehyde by Open Sandwich ELISA

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Abstract

Benzaldehyde (Bz) is a typical fragrant compound for peach-flavored beverages. In the food and beverage industries there is great demand for a sensitive and easy detection system of Bz in order to ensure product quality control and to avoid contamination. For the noncompetitive detection of Bz, we applied an open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) utilizing an antigen-dependent reassociation of antibody variable region fragments, VH and VL. We cloned the VH and VL genes of an anti-Bz monoclonal antibody, and the fragments were individually expressed and purified as a bacterial alkaline phosphatase (BAP)-conjugated form for VH and as a thioredoxine (Trx)-fused form for VL, respectively. Using these VH and VL fragments, we successfully constructed the OS-ELISA system for Bz detection. The Bz-induced formation of a trimolecular complex composed of VH-BAP/Bz/Trx-VL was readily detected by a dose-dependent increase in the BAP activity of the VH-fusion protein.

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Author notes

  1. Naoto Shirasu

    Present address: N.S. present address: Department of Biochemistry, Faculty of Medicine, Fukuoka University, 7-45-1 Nanakuma, Jonan, Fukuoka, 814-0180, Japan

Authors and Affiliations

  1. Department of Electronics, Graduate School of Information Science and Electrical Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka, 819-0935, Japan

    Naoto Shirasu, Takeshi Onodera & Kiyoshi Toko

  2. Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi, Fukuoka, 812-8581, Japan

    Kazutaka Nagatomo & Kiyoshi Matsumoto

  3. Department of Chemistry, Graduate School of Science, Kyushu University, 6-10-1 Hakozaki, Higashi, Fukuoka, 812-8581, Japan

    Yasuyuki Shimohigashi

Authors
  1. Naoto Shirasu
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  2. Takeshi Onodera
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  3. Kazutaka Nagatomo
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  4. Yasuyuki Shimohigashi
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  5. Kiyoshi Toko
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  6. Kiyoshi Matsumoto
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Correspondence to Naoto Shirasu.

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Shirasu, N., Onodera, T., Nagatomo, K. et al. Noncompetitive Immunodetection of Benzaldehyde by Open Sandwich ELISA. ANAL. SCI. 25, 1095–1100 (2009). https://doi.org/10.2116/analsci.25.1095

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  • DOI: https://doi.org/10.2116/analsci.25.1095

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