GLP-2融合蛋白的初步纯化

Primary Purification of Fusion Protein GLP-2

目的对包涵体的提取,洗涤等融合蛋白的获取条件进行研究,同时初步探索了融合蛋白的酸水解条件。方法包涵体用TEMFX洗涤1次,0.5%Triton洗涤4次,加入25%(v/v)的8 mol/L尿素变性溶解包涵体沉淀,再进行乙醇分级沉淀。结果所得融合蛋白纯度达到77.582%。按6%的比例加入60 mmol/L Hcl,50℃、48 h基本水解完全。结论应用本文方法对GLP融合蛋白初步纯化简便可行,为进一步研究奠定必要的基础。

Objective To research the access conditions of fusion protein including the extraction and washing inclusion body. And make a preliminary study on Acid hydrolysis conditions of fusion protein. Methods Inclusion body was washed by TEMFX once, and 0. 5% Triton 4 times. 25% 8 mol/L urea was added to denature and dissolve degeneration of inclusion body. At last alcohol was added to fractional precipitation. Results The purity of fusion protein is 77. 582%. The fusion protein is basic un-hydrolyzed by adding 60 mmol/L Hcl according to the proportion of 6%, 50 ℃, 48 h. Conclusion The method is simple and feasible and made essential basic preparing for research of next part.