Development of a simple and rapid method for producing non-fucosylated oligomannose containing antibodies with increased effector function.
Glycosylation in the Fc region of antibodies has been shown to play an important role in antibody function. In the current study, glycosylation of human monoclonal antibodies was metabolically modulated using a potent alpha-mannosidase I inhibitor, kifunensine, resulting in the production of antibodies with oligomannose-type N-glycans. Growing Chinese hamster ovary cells for 11 days in batch culture with a single treatment of kifunensine was sufficient to elicit this effect without any significant impact on cell viability or antibody production. Antibodies expressed in the presence of kifunensine at a concentration as low as 60 ng/mL contained mainly oligomannose-type glycans and demonstrated increased ADCC activity and affinity for FcgammaRIIIA, but reduced C1q binding. Although the kifunensine-mediated shift to oligomannose-type glycans could, in theory, result in rapid clearance of the antibody through increased mannose receptor binding, the serum levels of antibody in mice were not significantly altered up to 168 h following injection. The use of kifunensine provides a simple and rapid method for the production of antibodies with increased ADCC without the time-consuming need to re-engineer either the antibody molecule or the host cell line.
Zhou
Qun
Q
Genzyme Corporation, P.O. Box 9322, Framingham, Massachusetts 01701-9322, USA. qun.zhou@genzyme.com
Shankara
Srinivas
S
Roy
Andre
A
Qiu
Huawei
H
Estes
Scott
S
McVie-Wylie
Alison
A
Culm-Merdek
Kerry
K
Park
Anna
A
Pan
Clark
C
Edmunds
Tim
T
D000470:Alkaloids / Q000008:administration & dosage*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005643:Fucose / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D008358:Mannose / Q000737:chemistry* / Q000276:immunology*; D051379:Mice; D015202:Protein Engineering / Q000379:methods*
D000470:Alkaloids; D000911:Antibodies, Monoclonal; C065629:kifunensine; D005643:Fucose; D008358:Mannose