Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma |
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Authors: | K Tanaka S Hashida T Kohno Z Mohri Y Murakami E Ishikawa |
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Institution: | (1) Department of Biochemistry, Medical College of Miyazaki, 889-16 Kiyotake, Miyazaki, (Japan);(2) Research Laboratories, Research and Development Div., Sumitomo Pharmaceuticals Co. Ltd., Konohana, 554 Osaka, (Japan) |
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Abstract: | Summary A novel and sensitive noncompetitive enzyme immunoassay for kassinin is described. Kassinin was biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate. The biotinylated kassinin was trapped on antikassinin IgG-coated polystyrene balls and, after washing to eliminate other biotinylated substances, was eluted with HCl. The biotinylated kassinin eluted was reacted with anti-kassinin Fab'-peroxidase conjugate and trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of kassinin was 0.13 pg (0.1 fmol)/tube or 0.065 g/l of rat plasma, which was 750-fold or 15-fold lower than that for competitive radioimmunoassay. |
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Keywords: | Enzyme immunoassay peptide kassinin biotinylation |
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