title: Development of a Sandwich ELISA for Quantification of Gly m 4, a Soybean Allergen.
abstract: Gly m 4 is a key soybean allergen that causes allergic symptoms in the skin, gastrointestinal tract, or respiratory tract of sensitive individuals. To understand naturally variable levels of Gly m 4 among conventional soybean varieties, a sandwich ELISA was developed and validated using a mouse anti-Gly m 4 monoclonal antibody and a goat anti-Gly m 4 polyclonal antibody as capture and detection antibodies, respectively. The ELISA shows high specificity to Gly m 4 without any cross-reactivity to other soybean proteins and has a quantification range of 7.8-250 ng/mL using an Escherichia coli-produced recombinant Gly m 4, with 2.1 ng/mL being the limit of detection. Within the quantification range, the coefficients of variation of the intra-assay and interassay precision are less than 5 and 12%, respectively. Moreover, extraction efficiency and dilutional parallelism experiments were completed to demonstrate the assay is accurate. The validated assay was used to quantify Gly m 4 levels in 128 soybean samples from 24 conventional soybean varieties grown at 8 distinct geographical locations. There was a 13-fold difference between the least and greatest amounts of Gly m 4 concentrations among the samples, and the results demonstrate that the most significant sources of variability in Gly m 4 levels in the conventional varieties were related to location and variety.
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Tao
authors.initials: T
authors.lastname: Geng
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Kang
authors.initials: K
authors.lastname: Liu
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Ronald
authors.initials: R
authors.lastname: Frazier
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Lifang
authors.initials: L
authors.lastname: Shi
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Erin
authors.initials: E
authors.lastname: Bell
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Glenn
authors.affiliation: Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, United States.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Ward
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000276:immunology; D000818:Animals; D052179:Antigens, Plant / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006041:Goats; D051379:Mice; D008807:Mice, Inbred BALB C; D013025:Glycine max / Q000737:chemistry* / Q000276:immunology
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C530536:Gly m 4 allergen, Glycine max
keywords: ELISA; Gly m 4; allergy; natural variability; soybean