Impact of Thermal Processing on ELISA Detection of Peanut Allergens

This study examined the effect of heat treatment on the solubility of peanut proteins and compared the performances of two commercial ELISA kits (Veratox Quantitative Peanut Allergen Test and BioKits Peanut Assay Kit) for quantitation of peanut residues as affected by different heat treatments (moist and dry heat) and detection targets (mixture of proteins vs specific protein). Both laboratory-prepared and commercial peanut flour preparations were used for the evaluation. The two ELISA kits tended to underestimate the levels of protein in samples that were subjected to elevated heat, respectively, by more than 60- or 400-fold lower for the autoclaved samples and by as much as 70- or 2000-fold lower for the dark-roast commercial flour samples. The BioKits test, which employs antibodies specific to a heat labile protein (Ara h 1), in general exhibited a greater degree of underestimation. These results suggest that commercial ELISA kits may not be able to accurately determine the amount of proteins present in thermally processed foods due to changes in the solubility and immunoreactivity of the target proteins. Users need to be aware of such limitations before applying ELISA kits for evaluation of food allergen control programs.