Noncompetitive immunodetection of benzaldehyde by open sandwich ELISA

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

Research output: Contribution to journalArticlepeer-review

12 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, 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. 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

Fingerprint

Dive into the research topics of 'Noncompetitive immunodetection of benzaldehyde by open sandwich ELISA'. Together they form a unique fingerprint.

Cite this