Production of Recombinant Porcine Tumor Necrosis Factor Alpha in a Baculovirus Expression System.

Abstract

ABSTRACT. Porcine tumor necrosis factor alpha (TNF-α) was produced using a baculovirus system in Spodoptera frugiperda (SF21AE) cells. Cytotoxic activity was detected in supernatant of sonicated SF21AE cells infected with recombinant viruses. The recombinant protein was also demonstrated to be functionally active by its ability to cause apoptosis in Wehi 164 cells. Three distinct bands of 26, 17 and 14 KDa were revealed by Western blot analysis using anti-human TNF-α antibody. Moreover, the anti-human TNF-α antiserum significantly neutralized the cytotoxic activity of the supernatant of sonicated SF21AE cells infected with recombinant viruses. -KEY WORDS: baculovirus, porcine, tumor necrosis factor-alpha.

Biological Activities of Lipopolysaccharides Extracted from Porcine Vaccine Strains.

“…TNF-α was measured by cytotoxicity assay on Wehi 164 clone 28-4 cells, in the presence of 1 µg/ml actinomycin D (Dojindo), as described previously [16]. Recombinant mouse TNF-α (Genzyme, U.S.A.) was used as a standard TNF-α.…”