Journal
TRANSGENIC RESEARCH
Volume 18, Issue 3, Pages 467-482Publisher
SPRINGER
DOI: 10.1007/s11248-008-9240-1
Keywords
Monoclonal antibody; Plant; Tobacco; Culture cells; Proteolysis; Vacuole; Secretory pathway
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Funding
- European community (PHARMA-PLANTA integrated Project)
- Region Wallonne (DGTRE/SUBCELL)
- Inter-university Attraction Poles Program-Belgian Science Policy
- Belgian fund
- Fonds pour la Formation a la Recherche dans l'Industrie et dans l'Agriculture (Belgium)
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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.
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