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New Chromatography Data Have Been Reported by H. Liu and Co-Authors (Integrative refolding and purification of histidine-tagged protein by...

July 4, 2014



New Chromatography Data Have Been Reported by H. Liu and Co-Authors (Integrative refolding and purification of histidine-tagged protein by like-charge facilitated refolding and metal-chelate affinity adsorption)

By a News Reporter-Staff News Editor at Science Letter -- Investigators discuss new findings in Science. According to news reporting out of Tianjin, People's Republic of China, by NewsRx editors, research stated, "This work proposed an integrative method of protein refolding and purification by like-charged resin facilitated refolding and metal-chelate affinity adsorption. Hexahistidine-tagged enhanced green fluorescence protein (EGFP) was overexpressed in Escherichia coil as inclusion bodies (IBs), and then the protein was refolded and purified from urea-solubilized IBs by this method."

Our news journalists obtained a quote from the research, "A metal-chelating resin was fabricated by coupling iminodiacetic acid (IDA) to agarose gel (Sepharose FF). The anionic resin was used to facilitate the refolding of like-charged EGFP from IBs. After refolding, nickel ions were introduced for the affinity purification of the target protein by metal-chelating adsorption. It was found that the resin was effective in facilitating EGFP refolding. For 0.1 mg/mL EGFP IBs refolding, the fluorescence recovery (FR) by direct dilution was only 64%; addition of only 0.05 g/mL resin increased the FR to over 90%. Moreover, the FR increased with increasing resin concentration. Owning to the shielding effect of the oppositely charged impurities embedded in IBs on the surface charges of the IDA resin, more resin particles were required to exert an aggregation inhibition effect in the IBs protein refolding. Additionally, compared with direct-dilution refolding, inclusion of like-charged resins not only offered an enhanced FR of EGFP, but also bound some opposite-charged contaminant proteins, leading to a preliminary purification effect. Afterwards, the refolded EGFP was recovered by metal-chelating adsorption at an FR of 85% and purity of 93%."

According to the news editors, the research concluded: "This work has thus extended the like-charge facilitated protein refolding strategy to the integrative protein refolding and purification."

For more information on this research see: Integrative refolding and purification of histidine-tagged protein by like-charge facilitated refolding and metal-chelate affinity adsorption. Journal of Chromatography A, 2014;1344():59-65. Journal of Chromatography A can be contacted at: Elsevier Science Bv, PO Box 211, 1000 Ae Amsterdam, Netherlands. (Elsevier - www.elsevier.com; Journal of Chromatography A - www.elsevier.com/wps/product/cws_home/502688)

Our news journalists report that additional information may be obtained by contacting H. Liu, Collaborat Innovat Center Chem Sci & Engn Tianjin, Tianjin 300072, People's Republic of China. Additional authors for this research include W.J. Du, X.Y. Dong and Y. Sun (see also Science).

Keywords for this news article include: Asia, Tianjin, Science, People's Republic of China

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Source: Science Letter