TY - JOUR
T1 - Baculovirus-mediated large-scale expression and purification of a polyhistidine-tagged rubella virus capsid protein
AU - Schmidt, Michel
AU - Tuominen, Nina
AU - Johansson, Tove
AU - Weiss, Stefan
AU - Keinänen, Kari
AU - Oker-Blom, Christian
PY - 1998
Y1 - 1998
N2 - The capsid protein of rubella virus was produced in baculovirus-infectedSpodoptera frugiperdainsect cells, with a polyhistidine affinity tag at the carboxy terminus. The RV capsid recombinant protein was produced in a 10-liter bioreactor and purified, under nondenaturing conditions, using immobilized metal–ion affinity chromatography. Immunoblot analyses indicated that the purified recombinant protein was intact and migrated with the expected molecular weight. The final yield was 5 mg of purified protein per liter of cell culture. Surface plasmon resonance was used to investigate the antigenic potential of the histidine tagged capsid protein in an antigen–antibody interaction study. A specific interaction between the two proteins was shown. Our results suggest that this strategy should be useful in interaction studies of other virus-specific proteins and antibodies.
AB - The capsid protein of rubella virus was produced in baculovirus-infectedSpodoptera frugiperdainsect cells, with a polyhistidine affinity tag at the carboxy terminus. The RV capsid recombinant protein was produced in a 10-liter bioreactor and purified, under nondenaturing conditions, using immobilized metal–ion affinity chromatography. Immunoblot analyses indicated that the purified recombinant protein was intact and migrated with the expected molecular weight. The final yield was 5 mg of purified protein per liter of cell culture. Surface plasmon resonance was used to investigate the antigenic potential of the histidine tagged capsid protein in an antigen–antibody interaction study. A specific interaction between the two proteins was shown. Our results suggest that this strategy should be useful in interaction studies of other virus-specific proteins and antibodies.
U2 - 10.1006/prep.1997.0851
DO - 10.1006/prep.1997.0851
M3 - Article
SN - 1046-5928
VL - 12
SP - 323
EP - 330
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 3
ER -