title: High level expression on a chimeric anti-ganglioside GD2 antibody: genomic kappa sequences improve expression in COS and CHO cells.
abstract: We report a flexible strategy for the high level expression of a recombinant human monoclonal antibody (mAb) in Chinese hamster ovary (CHO) cells, initially using COS monkey kidney cell transfections to evaluate rapidly modifications to immunoglobulin (Ig) DNA constructs. Using sequential transfections with two amplifiable markers, we generated CHO cell lines and clones that secrete 80-110 micrograms/10(6) cells/24 hours of a mouse-human chimeric IgG1 kappa mAb. This cellular productivity is considerably greater than most murine hybridomas and transfected myelomas. Our data also demonstrate that genomic kappa sequences can improve mAb expression in COS and CHO cells. As a paradigm, we focused our expression studies on a human chimeric form of 3F8, a murine mAb that binds to ganglioside GD2 on neuroblastoma and melanoma tumor cells.
authors.affiliation: Genetics Institute, Cambridge, Massachusetts 02140.
authors.forename: L A
authors.initials: LA
authors.lastname: Fouser
authors.forename: S L
authors.initials: SL
authors.lastname: Swanberg
authors.forename: B Y
authors.initials: BY
authors.lastname: Lin
authors.forename: M
authors.initials: M
authors.lastname: Benedict
authors.forename: K
authors.initials: K
authors.lastname: Kelleher
authors.forename: D A
authors.initials: DA
authors.lastname: Cumming
authors.forename: G E
authors.initials: GE
authors.lastname: Riedel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016466:CHO Cells / Q000378:metabolism*; D006224:Cricetinae; D000939:Epitopes; D005732:Gangliosides; D015870:Gene Expression; D000882:Haplorhini; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G / Q000096:biosynthesis*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D005732:Gangliosides; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins