Fifteen hybridoma cell lines producing monoclonal antibodies (McAb) against recombinant human tumor necrosis factor alpha (rhTNFa) have been established by fusing SP 2/0 cells with spleen cells from a BALB/c mouse immunized with rhTNFa. Following J. M. Davis's Works, semi-solid medium was used for initial cloning. Five of them were studied further. Their main chromosome numbers range were 96 to 105, all of them were IgG1 subclass. The affinities of these McAbs were estimated to be 1.25 x 10(8) mol/L, 1.12 x 10(8) mol/L, 2.34 x 10(8) mol/L, 8.55 x 10(7) mol/L, 1.04 x 10(8) mol/L, respectively. Two groups of mice challenging with E. Coli (10(7) organisms), one group treated with 2 mg/kg anti-TNF monoclonal antibody, the other did not. There was a higher survival rate in treated group, the serum TNF level was significantly lower too, and the untreated mice had severe pathologic changes in viscera.