title: Homologous recombination in hybridoma cells: heavy chain chimeric antibody produced by gene targeting.
abstract: We demonstrate that murine myeloma cells can efficiently mediate homologous recombination. The murine myeloma cell line J558L was shown to appropriately recombine two transfected DNA molecules in approximately 30% of cells that received and integrated intact copies of both molecules. This activity was then exploited to direct major reconstructions of an endogenous locus within a hybridoma cell line. Production of antigen-specific chimeric heavy chain was achieved by targeting the human IgG1 heavy chain constant region (C gamma 1) exons to the genomic heavy chain locus of a hybridoma cell line secreting antibody specific for a human tumor-associated antigen. The frequency of productive genomic recombinations was approximately 1 in 200 transfectants, with accumulation of the chimeric protein reaching greater than 20 micrograms/ml in culture supernatants.
authors.affiliation: Oncogen, Seattle, WA 98121.
authors.forename: H P
authors.initials: HP
authors.lastname: Fell
authors.forename: S
authors.initials: S
authors.lastname: Yarnold
authors.forename: I
authors.initials: I
authors.lastname: Hellström
authors.forename: K E
authors.initials: KE
authors.lastname: Hellström
authors.forename: K R
authors.initials: KR
authors.lastname: Folger
mesh_terms: D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D015153:Blotting, Western; D002460:Cell Line; D002678:Chimera; D004797:Enzyme-Linked Immunosorbent Assay; D005803:Genes, Immunoglobulin; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D051379:Mice; D010954:Plasmacytoma; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection
chemical_list: D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains