Determination of glycinin in soybean and soybean products using a sandwich enzyme-linked immunosorbent assay

• Anti-glycinin monoclonal antibodies were produced based on soy-free mouse model.
• A sandwich ELISA is developed based on Mab 6F2 for detection of soybean glycinin.
• The assay shows high affinity and specificity in comparison with previous methods.
• We analysed the content of glycinin in 469 soybean seeds and five soybean products.

This study performs a sandwich ELISA for detection of trace amounts of glycinin in soybean products. We designed a soy-free mouse model to produce anti-glycinin monoclonal antibodies with high affinity and specificity. Using the monoclonal antibody as coating antibody, with the rabbit anti-glycinin polyclonal antibody as a detected antibody, the established sandwich ELISA showed high specificity for glycinin with minimum cross-reactions with other soy proteins. The practical working range of the determination was 3–200 ng/mL with detection limit of 1.63 ng/mL. The regaining of glycinin in spiked soybean samples were between 93.8% and 103.3% with relative standard deviation less than 8.3% (intra-day) and 10.5% (inter-day). The developed assay was used in analysing 469 soybean samples and five soybean products under different processing. The assay provides a specific and sensitive method for screening of glycinin and allows for further investigation into hypersensitive mechanisms to soybean proteins.