Large-scale purification of an antibody directed against hepatitis B surface antigen from transgenic tobacco plants.
The application of bioengineering to plants for production of biological products for human and animal use has expanded in recent years. The reasons for this expansion are several and include advances in the technology for novel production systems and the need for very large quantities of therapeutic proteins. The process of growing pharmaceutical proteins in plants, extracting, and purifying is a hard task considering the lack of available information concerning these topics. In this work, a recombinant murine monoclonal antibody specific for the hepatitis B surface antigen, expressed in stably transformed transgenic Nicotiana tabacum plants, was purified by means of a recombinant protein A Streamline chromatography as the main purification step. The antibody expression level varied with the age of the plants and the number of harvests from 40 to 15microg/ml and the maximum process yield was about 25mg of plantibody/kg of biomass. Protein A Streamline chromatography was successfully used in the purification process yielding a recovery of about 60% and a plantibody SDS-PAGE purity of over 90% but unexpectedly, previous clarification steps could not be totally avoided. The amino acid sequence recognized by this affinity purified plantibody was similar to its murine counterpart verifying the potentiality of plants to replace animals or bioreactors for large-scale production of this monoclonal antibody.
Valdés
Rodolfo
R
Plant Derived Antibody Production and Generation Group, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba.
Gómez
Leonardo
L
Padilla
Sigifredo
S
Brito
José
J
Reyes
Biunayki
B
Alvarez
Tatiana
T
Mendoza
Otto
O
Herrera
Orlando
O
Ferro
William
W
Pujol
Merardo
M
Leal
Vladimir
V
Linares
Marbelis
M
Hevia
Yasser
Y
García
Cristina
C
Milá
Lorely
L
García
Olga
O
Sánchez
Rafael
R
Acosta
Alfonso
A
Geada
Déborah
D
Paez
Rolando
R
Luis Vega
Jorge
J
Borroto
Carlos
C
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000378:metabolism*; D014026:Nicotiana / Q000378:metabolism*
D000906:Antibodies; D006514:Hepatitis B Surface Antigens