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J Chromatogr B Analyt Technol Biomed Life Sci. 2009 May 15;877(14-15):1561-7. doi: 10.1016/j.jchromb.2009.03.048. Epub 2009 Apr 7.

Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography.

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1
Department of Chemical and Natural Resources Engineering, Faculty of Engineering, Universiti Malaysia Pahang, Kuantan, Pahang, Malaysia.

Abstract

Nucleocapsid (N) protein of Nipah virus (NiV) is a potential serological marker used in the diagnosis of NiV infections. In this study, a rapid and efficient purification system, HisTrap 6 Fast Flow packed bed column was applied to purify recombinant histidine-tagged N protein of NiV from clarified feedstock. The optimizations of binding and elution conditions of N protein of NiV onto and from Nickel Sepharose 6 Fast Flow were investigated. The optimal binding was achieved at pH 7.5, superficial velocity of 1.25 cm/min. The bound N protein was successfully recovered by a stepwise elution with different concentration of imidazole (50, 150, 300 and 500 mM). The N protein of NiV was captured and eluted from an inlet N protein concentration of 0.4 mg/ml in a scale-up immobilized metal affinity chromatography (IMAC) packed bed column of Nickel Sepharose 6 Fast Flow with the optimized condition obtained from the method scouting. The purification of histidine-tagged N protein using IMAC packed bed column has resulted a 68.3% yield and a purification factor of 7.94.

PMID:
19395325
DOI:
10.1016/j.jchromb.2009.03.048
[Indexed for MEDLINE]
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