疏水相互作用层析分离重组人干扰素α2b

目的采用疏水相互作用层析分离重组人干扰素α2b,去除干扰素样品中的二聚体,得到高纯度的干扰素用于进一步的研究。方法首先采用阳离子交换层析纯化复性重组人干扰素α2b,去除了大部分的杂蛋白,然后采用疏水相互作用层析纯化重组人干扰素α2b,去除复性过程中产生的错误折叠体和二聚体,并考察盐浓度、pH值、流速和洗脱液中尿素对疏水相互作用层析纯化效果的影响。结果硫酸铵初始浓度1.2 mol/L、缓冲液pH值6.0、流速2.5 mL/min、洗脱液中添加尿素浓度为2 mol/L时疏水相互作用层析纯化效果最佳。最终得到的重组人干扰素α2b非还原型SDS-PAGE电泳均呈单一条带。结论确定了疏水层析纯化重组人干扰素α2b的最优条件,成功提取到具有高活性、高纯度的重组人干扰素α2b纯品。

Purification of recombinant human interferon alpha 2b by hydrophobic interaction chromatography

Objective To separate recombinant human interferon alpha 2b and remove the dimer by the hydrophobic interaction chromatography for the further study.Methods Firstly the cation exchange chromatography was employed to purify the refolded human interferon alpha 2b by removing of most contaminants.Then a further purification was carried out to remove the mis-folded recombinants in the renaturation process by hydrophobic interaction chromatography.Furthermore,the effects of the salt concentration,pH,flow rate and urea concentration on hydrophobic interaction chromatography were investigated.Results Under the optimal condition that was ammonium sulfate concentration 1.2 mol/L,pH 6.0,flow rate 2.5 mL/min and urea 2 mol/L the purified recombinant showed a single band on a non-reduced SDS-PAGE.Conclusion The optimal condition for puriying recombinant human interferon alpha 2b by the hydrophobic interaction chromatography was determined,and the recombinant with high activity and purity was successfully purified.