重组人胸腺素α原的制备及理化鉴定

Production and physical-chemical property determination of recombinant prothymosin α

目的建立rhproTα中试工艺并对rhproTα的理化性质和生物学活性进行鉴定。方法通过对培养条件的研究,建立了发酵培养工艺;通过对表达产物用酸沉、酚抽以及离子交换层析和反相层析等方法进行纯化;通过一系列分析方法对rhproTα的理化性质及生物活性进行鉴定。结果该因子在大肠杆菌中的表达水平为20%左右,纯度达到95%以上,测定其相对分子质量为11790、等电点为3.50,其N端序列、氨基酸组成、紫外分析等理化性质与理论值一致并具有较好的生物学活性。结论建立了rhproTα的中试工艺,完成了rhproTα的理化性质鉴定和生物活性测定,将有望进一步开发用于临床。

Objective To establish a fermentation technique for studying the physical-chemical properties and biological activity of prothymosin α （proTα）. Methods Through optimizing culture conditions, the fermentation technique was established. The purification of fermentation broth was processed by acid precipitation, phenol extraction, ion-exchange chromatography, and reversed-phase chromatography. The physical-chemical properties and biological activity of rhproTα were analyzed by a series of metheds. Results The expression level of rhproTα in E. coli reached about 20%. The purity of final products was more than 95%. At the same time, the physical-cbemical properties and biological activity of rhproTα were evaluated, which showed that the product was similar to the nature. Conclusion Through establishing the technique for fermentation of rhproTα and analyzing the physical-chemical properties and biological activity, the rhproTα could be expected to further development for clinical.