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

Michel Schmidt, Nina Tuominen, Tove Johansson, Stefan Weiss, Kari Keinänen, Christian Oker-Blom

Research output: Contribution to journalArticleScientificpeer-review

11 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)323-330
JournalProtein Expression and Purification
Volume12
Issue number3
DOIs
Publication statusPublished - 1998
MoE publication typeA1 Journal article-refereed

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Rubella virus
Baculoviridae
Capsid Proteins
Recombinant Proteins
Proteins
Surface Plasmon Resonance
Bioreactors
Affinity Chromatography
Histidine
Cell Culture Techniques
Molecular Weight
Viruses
Antibodies
polyhistidine

Cite this

Schmidt, Michel ; Tuominen, Nina ; Johansson, Tove ; Weiss, Stefan ; Keinänen, Kari ; Oker-Blom, Christian. / Baculovirus-mediated large-scale expression and purification of a polyhistidine-tagged rubella virus capsid protein. In: Protein Expression and Purification. 1998 ; Vol. 12, No. 3. pp. 323-330.
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abstract = "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.",
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Baculovirus-mediated large-scale expression and purification of a polyhistidine-tagged rubella virus capsid protein. / Schmidt, Michel; Tuominen, Nina; Johansson, Tove; Weiss, Stefan; Keinänen, Kari; Oker-Blom, Christian.

In: Protein Expression and Purification, Vol. 12, No. 3, 1998, p. 323-330.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Oker-Blom, Christian

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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.

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