Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture

• Published in: Biochemical and biophysical research communications Vol. 345; no. 2; pp. 602 - 607
• Main Authors: Girard, Loïc Stéphane, Fabis, Marzena Jolanta, Bastin, Maryse, Courtois, Didier, Pétiard, Vincent, Koprowski, Hilary
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 30.06.2006
• Subjects: Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Antibodies, Viral - biosynthesis; Antibodies, Viral - genetics; Antibody; Blotting, Western; Cell Culture Techniques; Culture Media - chemistry; Disposable plastic bioreactors; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation, Plant; Humans; Nerve Tissue Proteins - isolation & purification; Neutralization Tests; Nicotiana - cytology; Nicotiana - virology; Nicotiana tabacum; Plant cell culture; Plants, Genetically Modified - genetics; Rabies; Rabies virus; Rabies virus - immunology; Recombinant; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Xanthi; Disposable plastic bioreactors; Antibody; Xanthi; Plant cell culture; Rabies; Recombinant
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2006.03.219

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 μg/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.