Monoclonal Antibodies to Porcine Tumor Necrosis Factor Alpha: Development of an Enzyme-Linked Immunosorbent Assay

Abstract

Five hybridomas (4F4, 14H1, 9B4, 6E10, and 8G7) secreting antibodies to porcine tumor necrosis factor alpha (PTNF-α) were obtained from one fusion. Four of the 5 monoclonal antibodies (Mab) recognized recombinant PTNF-α (rPTNF-α) on western blot and were able to neutralize both rPTNF-α and native (released by porcine macrophages) PTNF (nPTNF-α, only 4F4, 14H1, and 9B4 tested for the neutralization of nPTNF-α) in vitro. A sandwich enzyme-linked immunosorbent assay (ELISA) for PTNF-α was developed using Mab 4F4 and purified rabbit polycloral antibodies against PTNF-α. The test detected PTNF-α concentrations as low as 400 pg/ml and did not cross react with native porcine TNF-β, recombinant human TNF-α, recombinant mouse TNF-β or native mouse TNF-α. The Mabs and the ELISA should be useful for assessing PTNF-α levels in swine serum during disease processes and possibly for alleviation of toxic effects of TNF-α.