Abstract
Recombinant DNA techniques were used to clone, construct and express the bifunctional molecule FV/IFN-γ. The FV/IFN-γ is a single-chain 42KD fusion protein expressed in E. coli under control of the strong T7 bacteriophage promoter in the expression vector pT7-7-FV-IFN-γ. The fused gene fragment FV-IFN-γ containing a single-chain anti-TAG72 FV gene fragment as well as the human recombinant cDNA fragment ofIFN-γ molecule. The renatured soluble form of FV/IFN-γ was purified from E. coli inclusion bodies using HTPT chromatography. The yield of this fusion protein was estimated at lOmg/L. Our data showed that the FV/IFN-γ molecule retained the TAG72 antigen-binding specificity and the IFN-γ activity as measured in ELISA, Western blotting and up-regulation of CEA expression by IFN-γ. Therefore, it may prove to be useful in targeting the biological effect of IFN-γ to tumor cells and stimulating its immune destruction.
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