Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system
✍ Scribed by Patrick H.C. van Berkel; Jolanda Gerritsen; Edwin van Voskuilen; Gerrard Perdok; Tom Vink; Jan G.J. van de Winkel; Paul W.H.I. Parren
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 440 KB
- Volume
- 105
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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.