离子交换色谱法分离纯化胸腺肽α_1的工艺研究

Studies on the Isolation and Purification of Thymosin α_1 by Ion-exchange Chromatography

通过对胸腺肽α1的分离纯化,提高胸腺肽α1的纯度,确定简单可行的纯化工艺技术。将QAE Sephadex A-25和D208、D301、717等阴离子交换介质进行静态吸附实验,确定最适离子交换介质;通过L(933)正交试验对流速、填料高度及进样浓度进行优化,确定最佳分离条件。以QAE Sephadex A-25作为填料介质吸附胸腺肽α1,在流速60 cm/h,填料高度50 cm,进样浓度8 mg/mL条件下,胸腺肽α1纯化效果理想,经HPLC测定,得到纯度达24.596 6 mg/mL的胸腺肽α1,质量分数达12.03%,比市场上销售的胸腺肽中Tα1含量(质量分数0.6%～1.0%)高出近10倍。

Through purifying thymosin α1and improve the purity of thymosin α1 in order to research a simple and feasible purification technology.The study chooses QAE Sephadex A-25 and D208,D301,717 ect.for static IV adsorption experiment through the anion exchange chromatography in order to determine the optimum ion exchange media;The optimal experiment conditions of the eluent,eluting style,flow rate,height of media are studied and choose L9（33） orthogonal experiment on optimizing the conditions of the velocity,height and the concentration.The study chooses HPLC and SDS-PAGE to identifying the thymosin α1.The experiment showed that the QAE Sephadex A-25 as a filler media adsorbing thymosin α1 makes the best effect,the velocity is 60 cm/h,packing height is 50 cm,the concentration is 8 mg/mL,then the purification results is satisfactory.Determined by the HPLC,the purity of thymosin α1is 24.596 6 mg/mL,the mass percent is 12.03 %.The method is accurate and feasible.The purity of thymosin α1is 10 times higher than the commercial thymosin α1 products（the mass percent is 0.6 % to 1.0 %）.