Anti - Tnf Mcab Development And Preliminary Application

TNF (Tumor Necrosis Factor) is a cytokine with anti -tumor activity was named by E. A. Carswell et al. in 1975. This polypeptide has been identifiedas one of such factors that has more bioactivities. The beneficial effects of TNF usually predominate with low level which produced host defenceimmunologically . In contrast, the high level of TNF will rampant tissue injury. Recombinant human TNF was established abroad in 1985, and its phase â…¡ clinical trial was studied in 1989. However, the results of several systemic clinical trial on tumor were q u i te disappointed. The administration of high-dose will increase the toxic side-effect. So the research focus were shifted to decreasing the TNF level in patients. McAb against TNF was such a antagonist that could lower TNF in serum.The purpose of this study was to produce the McAb against rhTNF, to establish ELISA test for healthy and patients, and to investigate the effect of TNF McAb on mice.Following J. M. Davis's work, semi-solid medium was used in this study for initial cloning. Briefly, after immunization and cell fusion, the initial cloning of hybridoma cells was carried on in this medium containing methylcellulose, IMDM and HAT . Five to seven days later, visible clones were removed to 96 -well plates for further subculture in a liquid medium containing DMEM, 15% FCS and HT . After screeningthe...