Noncompetitive immunodetection of benzaldehyde by open sandwich ELISA

Naoto Shirasu, Takeshi Onodera, Kazutaka Nagatomo, Yasuyuki Shimohigashi, Kiyoshi Toko, Kiyoshi Matsumoto

Research output: Contribution to journalArticle

11 Citations (Scopus)

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, V H and V L . We cloned the V H and V L genes of an anti-Bz monoclonal antibody, and the fragments were individually expressed and purified as a bacterial alkaline phosphatase (BAP)-conjugated form for V H and as a thioredoxine (Trx)-fused form for VL, respectively. Using these V H and V L fragments, we successfully constructed the OS-ELISA system for Bz detection. The Bz-induced formation of a trimolecular complex composed of V H- BAP/Bz/Trx-V L was readily detected by a dose-dependent increase in the BAP activity of the V H- fusion protein. 2009

Original languageEnglish
Pages (from-to)1095-1100
Number of pages6
Journalanalytical sciences
Volume25
Issue number9
DOIs
Publication statusPublished - Sep 2009

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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    Shirasu, N., Onodera, T., Nagatomo, K., Shimohigashi, Y., Toko, K., & Matsumoto, K. (2009). Noncompetitive immunodetection of benzaldehyde by open sandwich ELISA. analytical sciences, 25(9), 1095-1100. https://doi.org/10.2116/analsci.25.1095