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Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system
Authors:Patrick HC van Berkel  Jolanda Gerritsen  Edwin van Voskuilen  Gerrard Perdok  Tom Vink  Jan GJ van de Winkel  Paul WHI Parren
Institution:1. Genmab BV, PO BOX 85199, 3508 AD Utrecht, the Netherlands;2. telephone: 31302123337;3. fax: 31302123196
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 ~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 FcγRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells. Biotechnol. Bioeng. 2010; 105: 350–357. © 2009 Wiley Periodicals, Inc.
Keywords:human IgG  kifunensine  Fcγ  RIIIA  ADCC
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