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  • Title: Capture immunoassay for ruminant tumor necrosis factor-alpha: comparison with bioassay.
    Author: Ellis JA, Godson D, Campos M, Sileghem M, Babiuk LA.
    Journal: Vet Immunol Immunopathol; 1993 Jan; 35(3-4):289-300. PubMed ID: 8430498.
    Abstract:
    Monoclonal antibodies and IgG purified from rabbit polyclonal antiserum, raised against recombinant bovine tumor necrosis factor-alpha (TNF-alpha), have been employed in ELISA procedures to quantitate bovine TNF-alpha. These antibodies were potent in neutralizing the biological activity of recombinant as well as natural bovine TNF-alpha. 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-alpha 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-alpha 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.
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