Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture.
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.
Girard
Loïc Stéphane
LS
Thomas Jefferson University, 1020 Locust Street, Biotechnology Foundation, Philadelphia, PA 19107, USA.
Fabis
Marzena Jolanta
MJ
Bastin
Maryse
M
Courtois
Didier
D
Pétiard
Vincent
V
Koprowski
Hilary
H
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D003470:Culture Media; C029152:G-substrate; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins