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Abstract
A single-step purification method based on an anion exchange chromatography was developed to purify truncated Nipah virus glycoprotein (tNiVG) expressed in Spodoptera frugiperda 9 (Sf9) insect cells. The preferred buffer conditions to bind and elute tNiVG protein were 50 mM sodium carbonate with pH 9 and 50 mM sodium citrate with pH 5, respectively. The use of elution buffer without sodium chloride separated the tNiVG protein from the tightly bound major host proteins and subsequently avoided the desalting step as one of the further downstream processes. About 90% purity and 89% recovery of tNiVG protein were achieved with the developed purification method.
ACKNOWLEDGEMENTS
This study was supported by the Research University Grant Scheme (RUGS) from Universiti Putra Malaysia. S. Raksha was supported by the Malaysian Technical Cooperation Programme (MTCP) from the Ministry of Higher Education, Malaysia.
Notes
*A 3 mL of clarified cell homogenate containing 1.5 mg of tNiVG was loaded to the column and washed with 6 mL of respective binding buffer.
*A 3 mL of clarified cell homogenate containing 1.5 mg of tNiVG was loaded to the column. After washing with 10 CV of sodium carbonate (pH 9), elution was performed with 50 mM citrate buffer (pH 4 to 6).
*A 3 mL of clarified cell homogenate containing 1.8 mg of tNiVG was loaded to the SepFastTM Supor Q column. After washing with 10 CV of sodium carbonate (pH 9), elution was performed with 50 mM citrate buffer (pH 5).
*BEVs-Baculovirus-Expression Vector system, NA-Not available, CHO-Chinese Hamster Ovary, SIVmac-Simian Immunodeficiency virus, HIV-Human Immunodeficiency virus.
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