A novel enzyme-linked immunosorbent assay for quantification of soybean beta-conglycinin, a major soybean storage protein, in soybean and soybean food products.
Soybean (Glycine max L.) storage proteins are composed of two major components, beta-conglycinin and glycinin, corresponding to 7S and 11S globulins, respectively. Recently, soybean beta-conglycinin (7S globulin) has been reported to show beneficial functions in animals and human. To date, there is no method for the precise quantification of soybean beta-conglycinin in processed food products or soybean seeds. We report here a novel method for this purpose. At first, antibodies specifically reactive to the subunits of beta-conglycinin were prepared. And then, a direct enzyme-linked immunosorbent assay (ELISA) for the quantification of soybean beta-conglycinin in processed foods and seeds was developed. In this assay, the sample was treated with sodium dodecyl sulfate sample buffer followed by dilution with phosphate-buffered saline. The diluted samples were poured and coated onto an ELISA plate and reacted with rabbit anti-beta-conglycinin antibody and peroxidase-labeled anti-rabbit IgG. Finally, the bound peroxidase-labeled antibody was detected by colorimetric reaction. By using this system, it has been possible to measure soybean beta-conglycinin concentrations in several processed food products. In addition, this simple quantification ELISA system was demonstrated to be adaptable for the quantification of beta-conglycinin contents of various soybean cultivars.
Moriyama
Tatsuya
T
Laboratory of Molecular Function of Food, Graduate School of Agriculture, Kyoto University, Gokasho Uji, Kyoto 611-001 1, Japan. moriyama@kais.kyoto-u.ac.jp
Machidori
Mika
M
Ozasa
Sayaka
S
Maebuchi
Motohiro
M
Urade
Reiko
R
Takahashi
Koji
K
Ogawa
Tadashi
T
Maruyama
Nobuyuki
N
D000818:Animals; D000918:Antibody Specificity; D052179:Antigens, Plant; D015153:Blotting, Western; D003124:Colorimetry; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005916:Globulins / Q000032:analysis*; D011817:Rabbits; D055314:Seed Storage Proteins; D012639:Seeds / Q000737:chemistry; D045730:Soy Foods / Q000032:analysis*; D030262:Soybean Proteins / Q000032:analysis*; D013025:Glycine max / Q000737:chemistry*
D052179:Antigens, Plant; D005916:Globulins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max