This antibody-cytokine fusion protein was achieved by conjugating/fusing the Anti-gp240 antigen scFv to TNFα. It was expressed in CHO and purified with affinity chromatography. The immunocytokine retains the ability to bind the gp240 antigen as well as the biological activity of TNFα. The fusion protein was shown to enhance the in vitro killing of both TNF-sensitive and TNF-resistant human melanoma cells compared to native TNF. Further studies confirmed that the observed effects were antibody-mediated since competition with free antibody reduced the apparent cytotoxicity of the construct. This immunocytokine was designed for treating melanoma.
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