ELISA detection of hazelnut proteins: effect of protein glycation in the presence or absence of wheat proteins

Abstract

Hazelnuts are widely used in the food industry, especially confectionary foods. Nevertheless, these nuts contain several allergenic proteins that may be unexpectedly present as contaminants in various foods and may pose a serious threat to allergic consumers. The enzyme-linked immunosorbent assay (ELISA) is the preferred method to assess the level of hazelnut protein contamination. It is commonly used by both the food industry and enforcement agencies. Several ELISA kits are commercially available. However, protein detectability by ELISA may be affected by severe changes that proteins undergo during processing. The aim of this study is therefore to investigate the impact of processing on the ability to detect hazelnut protein by four commercial ELISA kits. Hazelnut proteins in the presence or absence of soluble wheat proteins were modified with glucose via the Maillard reaction. Changes in hazelnut proteins, such as the formation of protein-bound carbonyls, losses of reactive lysine residues and free amino groups, and severe aggregation dramatically affected the hazelnut protein detection by the commercial kits. The observed impact was highly dependent on the type of ELISA kit used.

Keywords:

• hazelnut allergy
• screening-ELISA
• detectability
• Maillard reaction
• reactive lysine
• protein bound carbonyls
• aggregation

Acknowledgements

The authors greatly appreciate the Belgian Science Policy (SD/AF/03A) for funding this research. The authors would like to thank the Neogen Corporation for kindly providing the Veratox for Hazelnut ELISA kits used in this study. Tatiana Cucu and Céline Platteau contributed equally to this paper.