An efficient approach for recombinant expression and purification of the viral capsid protein from beak and feather disease virus (BFDV) in Escherichia coli

Subir Sarker, Seyed Ali Ghorashi, Crystall M.D. Swarbrick, Yogesh B. Khandokar, Zainab Himiari, Jade K Forwood, Shane R. Raidal

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Structural insights into the biology of viruses such as beak and feather disease virus (BFDV) which do not replicate in cell cultures are increasingly reliant on recombinant methods for protein production and purification. Development of efficient methods for homogenous production of BFDV capsid protein is also essential for vaccine development and diagnostic purposes. In this study, two different plasmids (pMCSG21 and pMCSG24), three homologous BFDV capsid proteins, and two unique expression media (auto-induction and IPTG-induced expression) were trialled for over-expression of the BFDV in Escherichia coli. Over-expression was observed for all three recombinant targets of BFDV capsid protein using E. coli BL21 (DE3) Rosetta 2 cell lines under IPTG induction. These proteins could be purified using an optimized, two-step purification process using a buffer containing 20. mM N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), 500. mM NaCl and supplemented with 200. mM l-arginine at pH 10.5, to yield a soluble and stable protein of greater than 95% purity. The final concentration of purified protein was approximately fourteen-to-eighteen fold greater than that reported previously. Initial crystallization and X-ray diffraction confirm that the protein is structured in a manner consistent with icosahedral symmetry. Antigenicity of recombinant Cap was confirmed by immunoassay, verifying its validity for use in continued experimentation as a potential DNA vaccine, a reagent in diagnostic assays, and purified concentrated protein for structural and functional biology.

Original languageEnglish
Number of pages8
JournalJournal of Virological Methods
Publication statusPublished - 1 Apr 2015
Externally publishedYes


  • Antigenicity
  • BFDV
  • Crystallization
  • IPTG-induction
  • Protein over-expression
  • Viral capsid protein

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