Capture immunoassay for ruminant tumor necrosis factor-α: comparison with bioassay
Identifieur interne : 004366 ( Main/Exploration ); précédent : 004365; suivant : 004367Capture immunoassay for ruminant tumor necrosis factor-α: comparison with bioassay
Auteurs : John A. Ellis [Canada] ; Dale Godson [Canada] ; Manuel Campos [Canada] ; Maarten Sileghem [Kenya] ; Lorne A. Babiuk [Canada]Source :
- Veterinary Immunology and Immunopathology [ 0165-2427 ] ; 1993.
Abstract
Monoclonal antibodies and IgG purified from rabbit polyclonal antiserum, raised against recombinant bovine tumor necrosis factor-alpha (TNF-α), have been employed in ELISA procedures to quantitate bovine TNF-α. These antibodies were potent in neutralizing the biological activity of recombinant as well as natural bovine TNF-α. The monoclonal antibodies were used as capture antibodies and were either passively adsorbed or covalently linked to ELISA plates. Polyclonal rabbit anti-TNF IgG was used as the detecting antibody in combination with a biotinylated anti-rabbit serum and a streptavidin-horseradish peroxidase conjugate. The detection limit for recombinant TNF-α medium was 10 pg ml−1 and in bovine or ovine serum was 35 pg ml−1. A good correlation was found between the ELISA and the WEHI-164 Clone 13 biologic assay when TNF-α was measured in medium containing serum or in serum. This capture ELISA was also capable of detecting ovine, but not porcine. TNF in supernatants from cultures of lipopolysaccharide-stimulated pulmonary alveolar macrophages.
Url:
DOI: 10.1016/0165-2427(93)90040-B
Affiliations:
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These antibodies were potent in neutralizing the biological activity of recombinant as well as natural bovine TNF-α. The monoclonal antibodies were used as capture antibodies and were either passively adsorbed or covalently linked to ELISA plates. Polyclonal rabbit anti-TNF IgG was used as the detecting antibody in combination with a biotinylated anti-rabbit serum and a streptavidin-horseradish peroxidase conjugate. The detection limit for recombinant TNF-α medium was 10 pg ml−1 and in bovine or ovine serum was 35 pg ml−1. A good correlation was found between the ELISA and the WEHI-164 Clone 13 biologic assay when TNF-α was measured in medium containing serum or in serum. This capture ELISA was also capable of detecting ovine, but not porcine. 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