Monoclonal antibodies to human tumor necrosis factor α: In vitro and in vivo application

Abstract Three stable murine hybridoma cell lines, which secrete monoclonal antibodies (mAb) to human tumor necrosis factor α (TNFα), were established. None of the monoclonal antibodies cross-reacted with lymphotoxin, interleukin 2 (IL 2) or Interferon γ (IFNγ). The highly species-specific monoclonal antibody, designated as mAb 195, neutralizes the cytotoxic activity of human and chimpanzee TNFα. This antibody was further used during in vivo studies to neutralize human TNFα in a murine animal model. The mAb 114 is a weakly neutralizing antibody that binds to a different epitope of TNFα than mAb 195. mAb 114 shows a wide range of cross-reactivity with TNFα of the following species: dog, pig, cynomolgus, rhesus, baboon and chimpanzee. The mAb 199 binds to human TNFα, but does not neutralize the cytotoxic activity. The epitope recognized by this mAb is in close proximity to mAb 114. A reproducible, sensitive immunoassay for human TNF7α has been developed using the antibodies mAb 199 and mAb 195. The test is performed within 6 hr and detects TNF7α in serum samples, with a limit of detection of 5 to 10 pg/mL.