title: Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system.
abstract: Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes approximately 7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcgammaRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells.
authors.affiliation: Genmab BV, AD Utrecht, the Netherlands. p.vanberkel@genmab.com
authors.forename: Patrick H C
authors.initials: PH
authors.lastname: van Berkel
authors.forename: Jolanda
authors.initials: J
authors.lastname: Gerritsen
authors.forename: Edwin
authors.initials: E
authors.lastname: van Voskuilen
authors.forename: Gerrard
authors.initials: G
authors.lastname: Perdok
authors.forename: Tom
authors.initials: T
authors.lastname: Vink
authors.forename: Jan G J
authors.initials: JG
authors.lastname: van de Winkel
authors.forename: Paul W H I
authors.initials: PW
authors.lastname: Parren
mesh_terms: D000470:Alkaloids / Q000378:metabolism; D000920:Antibody-Dependent Cell Cytotoxicity; D001709:Biotechnology / Q000191:economics / Q000379:methods*; D002460:Cell Line; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D017452:Receptors, IgG / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013997:Time Factors
chemical_list: D000470:Alkaloids; C496124:FCGR3A protein, human; D007074:Immunoglobulin G; D017452:Receptors, IgG; D011994:Recombinant Proteins; C065629:kifunensine; D006026:Glycoside Hydrolases