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Single-chain antibody variable region-targeted interleukin-2 stimulates T cell killing of human colorectal carcinoma
1Saskatoon Cancer Center, Department of Microbiology, University of Saskatchewan, Canada.
Abstract:
Recombinant DNA techniques were used to clone, construct and express the bifunctional molecule FV/IL-2. The FV/IL-2 is a single-chain 37 kDa fusion protein expressed in Escherichia coli under control of the strong T7 bacteriophage promoter in the expression vector pT7-7-FV-IL-2. The fused gene fragment FV-IL-2 contains a single-chain anti-TAG72 FV gene fragment and the human recombinant cDNA fragment of the IL-2 molecule. The renatured soluble form of FV/IL-2 was purified from E. coli inclusion bodies using hydroxylapatite chromatography. The yield of this fusion protein was estimated at 2.0 mg/L. Our data showed that the FV/IL-2 molecule retained the TAG72 antigen-binding specificity and the IL-2 activity as measured in the standard T cell proliferation as well as cytotoxicity assays. Therefore, it may prove to be useful in targeting the biological effect of IL-2 to tumour cells and stimulating its immune destruction.