Development of an indirect competitive immunoassay for walnut protein component in food

Among food allergens, walnut is a frequent cause of adverse food reactions in allergic patients. In this study, the walnut allergen protein 2S albumin precursor (Jug r 1) cDNA was synthesised and cloned into the pGEX-6P-1 expression vector. The recombinant plasmids were transformed into Escherichia coli (E. coli) BL21(DE3) pLys for expression of protein Jug r 1. Polyclonal antibodies were prepared against the expressed purified Jug r 1 protein. An indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of walnut soluble proteins in processed foods was developed using the prepared polyclonal antibodies. The developed ELISA had a high specificity, walnut protein standard solution at 2.2 ng/mL [inhibition concentration (IC80) of the competitive test] was clearly identified by the ELISA. The mean recoveries ranged from 86% to 112%. The coefficient of variation (CV) for the 4 model foods was 6.4–8.7%.