Different subcellular localization and glycosylation for a functional antibody expressed in Nicotiana tabacum plants and suspension cells
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Genes encoding the heavy and light chains of LO-BM2, a therapeutic IgG antibody, were assembled in the tandem or inverted convergent orientation and expressed in Nicotiana tabacum plants and BY-2 suspension cells. The tandem construct allowed higher expression in both expression systems. A similar degradation pattern was observed for the secreted antibody recovered from the leaf intercellular fluid and BY-2 culture medium. Degradation increased with leaf age or culture time. Antibodies purified from leaf tissues and BY-2 cells were both functional. However, MS analysis of the N-glycosylation showed complex plant-type glycans to be the major type in the antibody purified from plants, whereas, oligomannosidic was the major glycosylation type in that purified from BY-2 cells. LO-BM2 was observed mainly in the endoplasmic reticulum of BY-2 cells while, in leaf cells, it was localized mostly to vesicles resembling prevacuolar compartments. These results and those from endoglycosidase H studies suggest that LO-BM2 is secreted from BY-2 cells more readily than from leaf cells where it accumulates in a post-Golgi compartment.
KeywordsMonoclonal antibody Plant Tobacco Culture cells Proteolysis Vacuole Secretory pathway
This work was supported financially by a grant from the European community (PHARMA-PLANTA integrated Project), the Région Wallonne (DGTRE/SUBCELL), the Inter-university Attraction Poles Program-Belgian Science Policy, and the Belgian fund for scientific research. BDM is the recipient of a fellowship from the Fonds pour la Formation à la Recherche dans l’Industrie et dans l’Agriculture (Belgium).
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