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Article type: Research Article
Authors: Xiang, Jima; * | Qi, Yumina | Cook, Dana | Moyana, Terenceb
Affiliations: [a] Saskatoon Cancer Center, Department of Microbiology, University of Saskatchewan, Saskatoon, Canada and Cleveland Clinic, Cleveland, OH, USA | [b] Saskatoon Cancer Center, Departments of Microbiology and Pathology, University of Saskatchewan, Saskatoon, Canada
Correspondence: [*] Correspondence and reprint requests to: Dr. Jim Xiang, Saskatoon Cancer Center, 20 Campus Drive, Saskatoon, Saskatchewan S7N 4H4, Canada.
Abstract: The construction, synthesis and expression of a genetically engineered bifunctional antibody/cytokine fusion protein is described. To target IFN-τ to tumor cells, recombinant antibody techniques were used to construct a RM4/IFN-τ fusion protein containing the chimeric anti-tumor F(ab′)2 (RM4) and the IFN-τ moiety. The recombinant cDNA of IFN-τ was linked to 3 prime end of the chimeric heavy-chain gene fragment (M4) containing the VH, the CHI and the hinge region to form the fused heavy-chain gene fragment M4-IFN-τ. Transfection of the M4-IFN-τ gene fragment into a myeloma derived cell line VKCK which produced the chimeric light-chain of the same antibody, allowed the transfectant secreting the bifunctional fusion protein RM4/IFN-τ. The RM4 /IFN-τ was purified by the affinity chromatography. Our data showed that the RM4/IFN-τ retained the TAG72 antigen-binding reactivity as well as the IFN-τ activity as measured in ELISA, FACS analysis of cell-surface TAG72 expression, immunohistochemical study, and up-regulation of cell-surface expression of CEA, HLA class I and class II antigens. Therefore, the bifunctional fusion protein RM4/IFN-τ may prove to be useful in targeting biological effects of the IFN-τ to tumor cells and in this way to stimulate the immune destruction of tumor cells.
Keywords: Fusion protein, anti-TAG72 antibody, IFN-τ
DOI: 10.3233/HAB-1996-7101
Journal: Human Antibodies, vol. 7, no. 1, pp. 2-10, 1996
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