Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system.
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.
van Berkel
Patrick H C
PH
Genmab BV, AD Utrecht, the Netherlands. p.vanberkel@genmab.com
Gerritsen
Jolanda
J
van Voskuilen
Edwin
E
Perdok
Gerrard
G
Vink
Tom
T
van de Winkel
Jan G J
JG
Parren
Paul W H I
PW
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
D000470:Alkaloids; C496124:FCGR3A protein, human; D007074:Immunoglobulin G; D017452:Receptors, IgG; D011994:Recombinant Proteins; C065629:kifunensine; D006026:Glycoside Hydrolases