TY - JOUR
T1 - Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture
AU - Girard, Loïc Stéphane
AU - Fabis, Marzena Jolanta
AU - Bastin, Maryse
AU - Courtois, Didier
AU - Pétiard, Vincent
AU - Koprowski, Hilary
PY - 2006/6/30
Y1 - 2006/6/30
N2 - 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.
AB - 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.
KW - Antibodies, Monoclonal/biosynthesis
KW - Antibodies, Viral/biosynthesis
KW - Blotting, Western
KW - Cell Culture Techniques
KW - Culture Media/chemistry
KW - Enzyme-Linked Immunosorbent Assay
KW - Gene Expression Regulation, Plant
KW - Humans
KW - Nerve Tissue Proteins/isolation & purification
KW - Neutralization Tests
KW - Plants, Genetically Modified/genetics
KW - Rabies virus/immunology
KW - Recombinant Proteins/biosynthesis
KW - Tobacco/cytology
U2 - 10.1016/j.bbrc.2006.03.219
DO - 10.1016/j.bbrc.2006.03.219
M3 - Article
C2 - 16690026
VL - 345
SP - 602
EP - 607
JO - Biochemistry Biophysics Research Communications
JF - Biochemistry Biophysics Research Communications
SN - 0006-291X
IS - 2
ER -