rapid high yield production of different glycoforms of ebola virus monoclonal antibody快速高产生产不同改变的埃博拉病毒单克隆抗体.pdfVIP

Rapid High Yield Production of Different Glycoforms of Ebola Virus Monoclonal Antibody 1 2 3 2 2 Alexandra Castilho , Natasha Bohorova , Josephine Grass , Ognian Bohorov , Larry Zeitlin , Kevin 2 3 1 Whaley , Friedrich Altmann , Herta Steinkellner * 1 Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria, 2 Mapp Biopharmaceutical, San Diego, California, United States of America, 3 Department of Chemistry, University of Natural Resources and Life Sciences, Vienna, Austria Abstract Background: Fc-glycosylation of monoclonal antibodies (mAbs) has profound implications on the Fc-mediated effector functions. Alteration of this glycosylation may affect the efficiency of an antibody. However, difficulties in the production of mAbs with homogeneous N-glycosylation profiles in sufficient amounts hamper investigations of the potential biological impact of different glycan residues. Methodology/Principal Findings: Here we set out to evaluate a transient plant viral based production system for the rapid generation of different glycoforms of a monoclonal antibody. Ebola virus mAb h-13F6 was generated using magnICON expression system in Nicotiana benthamiana, a plant species developed for commercial scale production of therapeutic proteins. h-13F6 was co-expressed with a series of modified mammalian enzymes involved in the processing of complex N- glycans. Using wild type (WT) plants and the glycosylation mutant DXTFT that synthesizes human like biantennary N- glycans with terminal N-acetylglucosamine on each branch (GnGn structures) as expression hosts we demo