This antibody-cytokine fusion protein was achieved by conjugating/fusing the Anti-EDB scFv to TNFα and IL12, where IL12 was fused to the amino-terminus of the L19 scFv with one TNFα monomer fused to the carboxy-terminus of the (IL12)-scFv. It was expressed in CHO and purified with affinity chromatography. The Immunocytokines retains the ability to bind the EDB as well as the biological activity of TNFα and IL12. This bi-antibody-cytokine fusion protein was active in vitro but it failed to localize in the tumors and exhibited limited therapeutic activity in immunocompetent 129sV mice bearing a sub cutaneous syngeneic F9 teratocarcinoma. This Immunocytokines was designed for treating Advanced solid tumors, Advanced pancreatic cancer, Metastatic melanoma.
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