Pooling of monoclonal antibodies for the rapid detection of commercially important finfish

Abstract

Fish are a valuable source of high-quality proteins, beneficial polyunsaturated fatty acids, and lipid-soluble vitamins in the human diet, but they also make up one of the “big eight” allergenic foods that are responsible for >90% of food allergy cases. A reliable and cost-effective analytical method is thus needed to detect fish protein in foods to protect fish-allergic individuals. Although immunoassays have been developed to detect many food allergens, there is no effective immunoassay capable of recognizing all common food fish species on the market. The selectivities of three monoclonal antibodies (mAbs), 8F5, 2G9 and 2F3, known to exhibit anti-finfish activity against cooked crude fish protein extract from red snapper, yellow fin tuna and swordfish, respectively, were each tested for 63 commercially important fish species using indirect enzyme-linked immunosorbent assays (iELISA). Based on the results, none of the mAbs showed cross-reactivity with all fish species tested; the cross reactivity patterns of these 3 mAbs complemented each other and none cross reacted with any non-fish samples. Pooling these mAbs (1:1:1) in an iELISA successfully detected a wide spectrum of commercially important fish species, including those subjected to various processing conditions. This pooled mAbs based iELISA exhibited high sensitivity, specificity and reproducibility, suggesting its potential utility as a qualitative assay for the regulatory enforcement of the food laws that protect fish sensitive consumers.