Single-chain antibody variable region-targeted interleukin-2 stimulates T cell killing of human colorectal carcinoma cells.
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.
Xiang
J
J
Saskatoon Cancer Center, Department of Microbiology, University of Saskatchewan, Canada.
Liu
E
E
Moyana
T
T
Qi
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D002277:Carcinoma / Q000276:immunology / Q000473:pathology*; D015179:Colorectal Neoplasms / Q000276:immunology / Q000473:pathology*; D003601:Cytotoxicity Tests, Immunologic; D004926:Escherichia coli; D005813:Genes, Synthetic; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D007376:Interleukin-2 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects* / Q000276:immunology; D014407:Tumor Cells, Cultured
D007135:Immunoglobulin Variable Region; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins