Development of a Specific Monoclonal Antibody for Grouper (Epinephelus guaza) Identification by an Indirect Enzyme-Linked Immunosorbent Assay

Abstract

Identification of fish species adulteration is important for consumer protection and the enforcement of food-labeling laws. A monoclonal antibody (MAb) generated against soluble muscle proteins from grouper (Epinephelus guaza) has been used in two indirect enzyme-linked immunosorbent assay (ELISA) formats (microtiter plates and immunostick tubes) for the rapid authentication of grouper fillets. The 3D12 MAb was produced with the use of the hybridoma technique and tested against several commonly consumed fish species by ELISA. The 3D12 MAb specifically reacted with grouper samples and could be useful for the discrimination of grouper among other, less-valued, fish species sold in the marketplace.

Determination of food authenticity by enzyme-linked immunosorbent assay (ELISA)

“…Furthermore, MAbs have been produced against fish muscle proteins being possible the identification of the red snapper (Huang, Marshall, Kao, Otwell, & Wei, 1995) or the discrimination of raw and cooked grouper among other less valued fish species (Asensio et al, 2003a(Asensio et al, , 2003b.…”

Detection of grouper mislabelling in the fish market by an immunostick colorimetric ELISA assay

“…In a previous work, we reported the development of a monoclonal antibody specific to grouper (3D12 MAb) (Asensio et al, 2003). Since the new fish fillet markets provide cheap fish species not marketed before, such as meagre, Nile tilapia, sutchi catfish, amber jack, striped mullet, pink cusk-eel and kingklip, among others, which potentially could be used as substitutes for grouper, we have evaluated the specificity of the 3D12 MAb against these and other fish species (see Materials and methods section) by using an immunostick colorimetric ELISA.…”

“…On the basis of this information, this paper reports the application of an immunostick ELISA for faster detection of inappropriately labelled grouper fillets in the marketplace: an indirect ELISA assay that uses a monoclonal antibody specific to grouper muscle proteins (Asensio et al, 2003). The simplicity of the procedure and the short time required for the analysis make it suitable as a tool of implementation of the traceability and as a field test for fish processors and inspectors.…”

PCR-based methodology for the authentication of grouper (Epinephelus marginatus) in commercial fish fillets

“…Numerous analytical techniques which rely on proteins analysis have been developed for fish species identification: electrophoretic, chromatographic and immunological techniques (Asensio et al, 2003;Ataman, Celik, & Rehbein, 2006;Horstkotte & Rehbein, 2003). However, these methods are limited to discriminate, especially if they are antibody-based methods.…”