【摘要】 采用离子交换法分离纯化,对影响分离的多种因素包括吸附动力学、pH值、缓冲液和树脂的种类进行了深入讨论。结果发现,采用SPsepharoseFF树脂、pH4.4的醋酸缓冲液分离效果较好,经过方案优化,在pH4.4醋酸缓冲液平衡、盐梯度洗脱条件下纯化TP5,HPLC和质谱分析表明,一步分离纯度可达98%以上。该法可以满足大规模制备的要求。更多还原

【Abstract】 Nowadays, reverse phase liquid chromatography (RPLC) is the usual way to purify the peptide. But the efficiency of this method is low and the cost is high, and it is also limited to small scales. In this paper, we use the ion-exchange chromatography and discuss several factors that will influence the process of separation, such as adsorption dynamics, pH, buffers and variety of resins. The experiments show that when using SP sepharose FF resin and pH 4.4 acetic buffers, we will get good results of separation. After optimizing the experiments’ conditions, we found that under pH 4.4 acetic buffer binding and salt elution, the purity of TP5 which was analyzed with high pressure liquid chromatography (HPLC) and mass spectrum (MS) could reach 98% or more in one step. This method can also satisfy the requirements of large-scale preparation.更多还原