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Purification of long helical capsid of newcastle disease virus from Escherichia coli using anion exchange chromatography

Authors

  • Chee Fai Yap,

    1. Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia
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  • Wen Siang Tan,

    1. Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia
    2. Dept. of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia
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  • Chin Chin Sieo,

    1. Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia
    2. Dept. of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia
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  • Beng Ti Tey

    Corresponding author
    1. Chemical Engineering Discipline, School of Engineering, Monash University, Sunway Campus, Jalan Lagoon Selatan, Selangor, Malaysia
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Abstract

NPΔc375 is a truncated version of the nucleocapsid protein of Newcastle disease virus (NDV) which self-assembles into a long helical structure. A packed bed anion exchange chromatography (PB-AEC), SepFastTM Supor Q pre-packed column, was used to purify NPΔc375 from clarified feedstock. This PB-AEC column adsorbed 76.2% of NPΔc375 from the clarified feedstock. About 67.5% of the adsorbed NPΔc375 was successfully eluted from the column by applying 50 mM Tris-HCl elution buffer supplemented with 0.5 M NaCl at pH 7. Thus, a recovery yield of 51.4% with a purity of 76.7% which corresponds to a purification factor of 6.5 was achieved in this PB-AEC operation. Electron microscopic analysis revealed that the helical structure of the NPΔc375 purified by SepFastTM Supor Q pre-packed column was as long as 490 nm and 22–24 nm in diameter. The antigenicity of the purified NPΔc375 was confirmed by enzyme-linked immunosorbent assay. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 564–567, 2013

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