title: Determination of glycinin in soybean and soybean products using a sandwich enzyme-linked immunosorbent assay.
abstract: 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.
authors.affiliation: State Key Laboratory of Animal Nutrition, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, PR China.
authors.forename: Jingshu
authors.initials: J
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Animal Nutrition, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, PR China.
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Animal Nutrition, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, PR China.
authors.forename: Peixia
authors.initials: P
authors.lastname: Song
authors.affiliation: State Key Laboratory of Animal Nutrition, China Agricultural University, No. 2 Yuanmingyuan West Road, Beijing 100193, PR China. Electronic address: maxi@cau.edu.cn.
authors.forename: Xi
authors.initials: X
authors.lastname: Ma
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005916:Globulins / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D030262:Soybean Proteins / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D005916:Globulins; D030262:Soybean Proteins; C011474:glycinin
keywords: ELISA; Glycinin; Monoclonal antibody; Polyclonal antibody; Soy-free diet; Soybean