Development of a sensitive sandwich ELISA with broad species specificity for improved fish allergen detection

Ji Liang; Stephen L Taylor; Joseph Baumert; N Alice Lee

doi:10.1016/j.foodchem.2022.133656

Development of a sensitive sandwich ELISA with broad species specificity for improved fish allergen detection

Food Chem. 2022 Dec 1:396:133656. doi: 10.1016/j.foodchem.2022.133656. Epub 2022 Jul 9.

Authors

Ji Liang¹, Stephen L Taylor², Joseph Baumert², N Alice Lee³

Affiliations

¹ ARC Training Centre for Advanced Technologies in Food Manufacture, School of Chemical Engineering, University of New South Wales, New South Wales, Australia.
² Food Allergy Research and Resource Program, Department of Food Science and Technology, University of Nebraska-Lincoln, Nebraska, United States.
³ ARC Training Centre for Advanced Technologies in Food Manufacture, School of Chemical Engineering, University of New South Wales, New South Wales, Australia. Electronic address: alice.lee@unsw.edu.au.

PMID: 35839724
DOI: 10.1016/j.foodchem.2022.133656

Abstract

A sensitive Enzyme-linked Immunosorbent Assay (ELISA) with improved broad species specificity was developed for the detection of southern hemisphere fish residues in processed foods. The polyclonal antibodies were raised against parvalbumins from 13 fish species representing 7 fish orders selected for their molecular diversity and immunoreactivity profile. The optimized ELISA-2 (based on the rabbit capture antibody (RB#4) - sheep detection antibody (S2#4) pair) displayed an improved detection limit of 0.6 μg/L (3.7 μg of /kg). Our immunoreactivity-directed species selection approach in the strategized antibody production significantly improved the detection of no or weakly immunoreactive fish species previously not detected immunochemically. Of 37 commercially important fish species tested, the ELISA-2 could detect 28 fish species (76%). The optimized sample extraction with a buffer additive achieved good protein recoveries of 87.2 - 117.3% (within the AOAC recommended range). The ELISA-2 was able to detect fish residues in five highly processed food products.

Keywords: Allergen detection; Broad specificity; ELISA; Parvalbumin (PAV); Southern hemisphere.

MeSH terms

Allergens*
Animals
Antibodies
Enzyme-Linked Immunosorbent Assay
Fishes*
Parvalbumins
Rabbits
Sensitivity and Specificity
Sheep
Species Specificity

Substances

Allergens
Antibodies
Parvalbumins