Purification of long helical capsid of Newcastle disease virus from Escherichia coli using anion exchange chromatography
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 c...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Institute of Chemical Engineers
2013
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| Online Access: | http://psasir.upm.edu.my/id/eprint/28069/1/28069.pdf http://psasir.upm.edu.my/id/eprint/28069/ http://onlinelibrary.wiley.com/wol1/doi/10.1002/btpr.1697/abstract |
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| Institution: | Universiti Putra Malaysia |
| Language: | English |
| Summary: | 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. |
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