This antibody-cytokine fusion protein was achieved by conjugating/fusing the Anti-EDB scFv to IFNγ. It was expressed in CHO and purified with affinity chromatography. The immunocytokine retains the ability to bind the EDB as well as the biological activity of IFNγ. The scFv (L19) directed against the EDB domain of fibronectin was fused to the N-terminus of a cysteine-free mutant of mouse IFN-γ. The resulting fusion protein was capable of targeting new blood vessels in solid tumors, and the targeting efficiency was strikingly increased in tumor-bearing knockout mice lacking the interferon-γ receptor. ScFv (L19)-interferon-γ displayed a strong antitumor effect in both subcutaneous and metastatic murine F9 teratocarcinomas, but was not efficacious as single agent when used to treat C51 and CT26 tumors. This immunocytokine was designed for treating Advanced solid tumors, Advanced pancreatic cancer, Metastatic melanoma.
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