Development of an egg detection sandwich ELISA kit using monoclonal antibodies

Abstract

We developed a novel ELISA kit for egg detection in accordance with the official Japanese method for extraction (official extraction method), which is used for monitoring the appropriateness of food product labeling in Japan. The monoclonal antibody used in the kit was prepared using reduced and carboxymethylated chicken ovalbumin as an immunizing antigen. The newly developed ELISA kit detected egg white solubilized in 2- mercaptoethanoland sodium dodecylsul fate, which is used in the official extraction method. The detectable range of egg protein concentrations was 0.8-50.0 ng/mL (equivalent to 0.3-20.0 μg/g food). Cross-reactivity with both duck and Japanese quailegg was observed, but not with other food ingredients. Spike recovery tests using model processed foods showed that recovery ranged from 73.6 %.118.0 %, and intra- and inter-assay coefficients of variation were less than 6.0% and 9.1 %, respectively. Based on the official extraction method for quantitative analysis of egg in specified food ingredients, these results suggest that a simple, monoclonal antibody-based ELISA kit is as effective as the conventionalmethod for egg detection. (Received May 31, 2013 ; Accepted Sep. 26, 2013). © Copyright 2014, Japanese Society for Food Science and Technology.