title: A genetically engineered single-gene-encoded anti-TAG72 chimeric antibody secreted from myeloma cells.
abstract: SP2/0Ag14 murine myeloma cells transfected with the expression vector mpSV2neo-EP-FV-CH2-3-PA containing the single gene FV-CH2-3 secreted a single-gene-encoded chimeric antibody molecule FV/M4. This single-chain protein consisted of the heavy- and light-chain variable (VH and VL) domains covalently joined through a flexible linker peptide, while the carboxyl end of VL domain was connected to the amino terminus of hinge region of the ccM4 heavy-chain. Our data showed that the FV/M4 retained both its immunoreactivity for tumor-associated TAG72 antigen and its cytolytic activity to tumor cells as did the parental ccM4 antibody. Therefore, this single-gene-construct approach circumvents inefficiencies inherent in delivering two genes into a mammalian cell for assembly of a functional chimeric antibody and provides an alternative for construction of chimeric antibodies. It is particularly attractive for ex vivo transfection of cells from patients for certain gene-therapy modalities not only for cancer but also for a range of diseases in which immunotherapeutic approaches are used.
authors.affiliation: Saskatoon Cancer Center, Department of Microbiology, University of Saskatchewan, Canada.
authors.forename: Y
authors.initials: Y
authors.lastname: Qi
authors.forename: J
authors.initials: J
authors.lastname: Xiang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D000917:Antibody Formation; D000920:Antibody-Dependent Cell Cytotoxicity; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000276:immunology; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D004274:DNA, Recombinant; D000939:Epitopes; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins; C047052:tumor-associated antigen 72