Investigation of allergen proteins in five tomato cultivars

Abstract

Investigation of putative allergens from tomato berries is challenging as differences between human serum IgE specificity and reactivity as well as the non-specific binding of the primary and secondary antibodies often cause difficulties. In this study five tomato cultivars were investigated to evaluate their potential allergenicity in Hungarian tomato sensitive patients. The major allergens proved to be low molecular weight proteins, but several previously described allergens could be identified as well using IgE-Western blotting. IgE binding to cross-reactive carbohydrate determinants (CCDs) was ruled out through the use of CCD inhibitor, but non-specific binding of the secondary antibody remained an issue. IgE binding activity of a purified, immunoblot positive protein (band at 40 kDa), and non-specific binding of the secondary antibody to the same protein, was demonstrated with an Optical Waveguide Lightmode Spectroscopy (OWLS) based immunosensor. LC-ESI-MS/MS analysis showed this protein is an as-yet undescribed vicilin-type putative allergen.