Recombinant bifunctional molecule FV/IFN-gamma possesses the anti-tumor FV as well as the gamma interferon activities

• Published in: Cancer biotherapy Vol. 8; no. 4; p. 327
• Main Authors: Xiang, J, Qi, Y, Luo, X, Liu, E
• Format: Journal Article
• Language: English
• Published: United States 1993
• Subjects: Antigens, Neoplasm - immunology; Base Sequence; Carcinoembryonic Antigen - analysis; Escherichia coli - genetics; Genetic Vectors; Glycoproteins - immunology; Humans; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - chemistry; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - pharmacology; Interferon-gamma - biosynthesis; Interferon-gamma - chemistry; Interferon-gamma - genetics; Interferon-gamma - pharmacology; Molecular Sequence Data; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - pharmacology; Recombinant Proteins; Tumor Cells, Cultured
• ISSN: 1062-8401
• DOI: 10.1089/cbr.1993.8.327

Recombinant DNA techniques were used to clone, construct and express the bifunctional molecule FV/IFN-gamma. The FV/IFN-gamma 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-gamma. The fused gene fragment FV-IFN-gamma containing a single-chain anti-TAG72 FV gene fragment as well as the human recombinant cDNA fragment of IFN-gamma molecule. The renatured soluble form of FV/IFN-gamma was purified from E. coli inclusion bodies using HTPT chromatography. The yield of this fusion protein was estimated at 10mg/L. Our data showed that the FV/IFN-gamma molecule retained the TAG72 antigen-binding specificity and the IFN-gamma activity as measured in ELISA, Western blotting and up-regulation of CEA expression by IFN-gamma. Therefore, it may prove to be useful in targeting the biological effect of IFN-gamma to tumor cells and stimulating its immune destruction.