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Glycinin ELISA for determination of Soybean products

hits:1871     Date:09/20/17
Soybean glycinin is a major food allergen causing anaphylaxis. A sensitive detection method for glycinin is needed to evaluate soybean allergies in food and feed products. In the present study, monoclonal antibodies (Mabs) against glycinin were prepared using purified glycinin as the immunogen. The generated Mabs, named 3B2 and 4B2, were identified as being IgG2b and IgG2a iso-types respectively, and exhibited high specificity to glycinin. Then we developed a competitive ELISA based on Mab 4B2 to measure glycinin which showed an IC50 value of 1.7 ng/mL with a detection limit of 0.3 ng/mL, and the linear portion of the curve was 0.3鈥11.2 ng/mL. Recovery tests indicated that the competitive ELISA based on Mab 4B2 gave reliable reproducibility. The produced Mab 4B2 and the developed ELISA could provide a valuable tool for sensitive determination of glycinin and for future studies conducted to reveal the mechanism of how glycinin functions in anaphylaxis.
Using 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鈥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.
Unibiotest research Glycinin ELISA Kit, detect Limit is 90 ng /mL (ppb) ,is a competitive enzyme immunoassay for the quantitative analysis of Glycinin in soybean, feed, soybean meal, fermented soybean meal, extruded soybean, soy protein isolate, soy protein concentrate, samples.
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