Baculovirus-mediated large-scale expression and purification of a polyhistidine-tagged rubella virus capsid protein.
The capsid protein of rubella virus was produced in baculovirus-infected Spodoptera frugiperda insect 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.
Schmidt
M
M
VTT Biotechnology and Food Research, Espoo, FIN-02044 VTT, Finland.
Tuominen
N
N
Johansson
T
T
Weiss
S A
SA
Keinänen
K
K
Oker-Blom
C
C
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000502:physiology; D019149:Bioreactors; D002213:Capsid / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D006639:Histidine / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012412:Rubella virus / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000235:genetics / Q000821:virology; D013997:Time Factors
D007093:Imidazoles; D007106:Immune Sera; D010455:Peptides; D011994:Recombinant Proteins; C033223:polyhistidine; D006639:Histidine