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Expression, purification, and characterization of VP2 capsid protein of canine parvovirus in Escherichia coli

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Abstract

The capsid protein VP2 of canine parvovirus (CPV) was expressed in Escherichia coli using pMAL-c2x vector. After codon optimization, the mutant resulted in high-level expression of fusion protein. A simple procedure was used to purify fusion protein and remove endotoxin from fusion protein preparations. The fusion protein was antigenical similar to the native capsid protein as Western-blot assay performed with polyclonal antibodies obtained from dogs vaccinated with CPV. The immunogenicity of fusion protein was demonstrated in a vaccination experiment conducted with rabbits subcutaneously immunized. Rabbits vaccinated with fusion protein developed high titers of virus-specific antibodies response that neutralized CPV in vitro.

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Abbreviations

IPTG:

Isopropyl-β- d-thiogalactopyranoside

MBP:

Maltose-binding protein

OPD:

Orthophenylenediamine

CPV:

Canine parvovirus

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Acknowledgments

The authors thank the assistance of researchers involved in the development of this work. This study was funded by the National High Technology Research & Development Program of China (863 Program) (2002AA624010) and Key Science & Technology Program of Guangdong Province of China (2003A3050402).

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Correspondence to Fanquan Zeng.

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Zeng, F., Yeung, W., Lu, Y. et al. Expression, purification, and characterization of VP2 capsid protein of canine parvovirus in Escherichia coli . World J Microbiol Biotechnol 24, 457–463 (2008). https://doi.org/10.1007/s11274-007-9493-5

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Keywords

  • Canine parvovirus
  • Expression
  • Optimization
  • pMAL-c2x vector
  • VP2 protein