title: A recombinant cell line expressing a form of the Y13-259 anti-p21ras antibody which binds protein A and may be produced as ascites.
abstract: We have recently reported the molecular cloning of the immunoglobulin genes encoding the variable regions of the rat anti-p21ras antibody, Y13-259. These genes were reassembled into expression vectors supplying DNA sequences encoding human gamma 1 and kappa constant domains, as well as the leader sequence for antibody secretion, thus yielding Hu-Y13-259, a secretory anti-p21ras antibody containing human constant regions. We now report the creation of a recombinant cell line, NS0/Hu-Y13-259/B6, secreting high levels of the Hu-Y13-259 Ig. The antigen specificity of this recombinant antibody was demonstrated to be identical to that of the parental Y13-259, i.e. the amino acid sequence 60-76 of the p21ras protein. Unlike the parental cell line, the recombinant cells could be grown as ascites in mice, allowing the production of large quantities of the protein A-binding Hu-Y13-259 antibody.
authors.affiliation: CNR Institute of Neurobiology, Rome, Italy.
authors.forename: T M
authors.initials: TM
authors.lastname: Werge
authors.forename: A
authors.initials: A
authors.lastname: Bradbury
authors.forename: A
authors.initials: A
authors.lastname: Di Luzio
authors.forename: A
authors.initials: A
authors.lastname: Cattaneo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism*; D000918:Antibody Specificity; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D008070:Lipopolysaccharides / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000378:metabolism; D010957:Plasmids; D016283:Proto-Oncogene Proteins p21(ras) / Q000276:immunology; D051381:Rats; D013205:Staphylococcal Protein A / Q000378:metabolism*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides; D013205:Staphylococcal Protein A; C050016:lipoarabinomannan; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)