Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture |
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Authors: | Girard Loïc Stéphane Fabis Marzena Jolanta Bastin Maryse Courtois Didier Pétiard Vincent Koprowski Hilary |
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Affiliation: | Thomas Jefferson University, 1020 Locust Street, Biotechnology Foundation, Philadelphia, PA 19107, USA. |
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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. |
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Keywords: | Antibody Disposable plastic bioreactors Plant cell culture Rabies Recombinant Xanthi |
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