Baculovirus-mediated large-scale expression and purification of a polyhistidine-tagged rubella virus capsid protein

Protein Expression and Purification
M SchmidtC Oker-Blom

Abstract

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.

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Citations

Feb 26, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xindu Geng, Lili Wang
Dec 28, 1999·Journal of Molecular Recognition : JMR·D G Myszka
Mar 10, 2001·Protein Expression and Purification·H TuominenA Goldman
Dec 20, 1999·Journal of Biotechnology·A OrellanaC Oker-Blom

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