title: Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture.
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.
authors.affiliation: Thomas Jefferson University, 1020 Locust Street, Biotechnology Foundation, Philadelphia, PA 19107, USA.
authors.forename: Loïc Stéphane
authors.initials: LS
authors.lastname: Girard
authors.forename: Marzena Jolanta
authors.initials: MJ
authors.lastname: Fabis
authors.forename: Maryse
authors.initials: M
authors.lastname: Bastin
authors.forename: Didier
authors.initials: D
authors.lastname: Courtois
authors.forename: Vincent
authors.initials: V
authors.lastname: Pétiard
authors.forename: Hilary
authors.initials: H
authors.lastname: Koprowski
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000235:genetics; D015153:Blotting, Western; D018929:Cell Culture Techniques; D003470:Culture Media / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant; D006801:Humans; D009419:Nerve Tissue Proteins / Q000302:isolation & purification; D009500:Neutralization Tests; D030821:Plants, Genetically Modified / Q000235:genetics*; D011820:Rabies virus / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014026:Nicotiana / Q000166:cytology* / Q000821:virology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D003470:Culture Media; C029152:G-substrate; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins