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Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture
Authors:Girard Loïc Stéphane  Fabis Marzena Jolanta  Bastin Maryse  Courtois Didier  Pétiard Vincent  Koprowski Hilary
Institution:Thomas Jefferson University, 1020 Locust Street, Biotechnology Foundation, Philadelphia, PA 19107, USA.
Abstract:A Nicotiana tabacum cv. Xanthi cell culture was initiated from a transgenic plant expressing a human anti-rabies virus monoclonal antibody. Within 3 months, plant cell suspension cultures were established and recombinant protein expression was examined. The antibody was stably produced during culture growth. ELISA, protein G purification, Western blotting, and neutralization assay confirmed that the antibody was fully processed, with association of light and heavy-chains, and that it was able to bind and neutralize rabies virus. Quantification of antibody production in plant cell suspension culture revealed 30 microg/g of cell dry weight for the highest-producing culture (0.5 mg/L), 3 times higher than from the original transgenic plant. The same production level was observed 3 months after cell culture initiation. Plant cell suspension cultures were successfully grown in a new disposable plastic bioreactor, with a growth rate and production level similar to that of cultures in Erlenmeyer flasks.
Keywords:Antibody  Disposable plastic bioreactors  Plant cell culture  Rabies  Recombinant  Xanthi
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