重组人干扰素-γ的凝胶色谱分离及其变性剂对分离结果的影响

Separation of Recombinant Human Interferon Gamma on Gel Exclusion Chromatography and the Effect of Denaturating Agents on Separation Results

使用十二烷基硫酸钠、盐酸胍和尿素三种不同的变性剂溶解基因重组人干扰素-Gamma(r-IFN-γ)包含体,然后进行凝胶色谱分离。使用活性、电泳和高效疏水作用色谱三种方法,比较了三种变性剂对分离结果的影响。结果为:尿素溶解包含体后r-IFN-γ的分子量与杂质相差较大,盐酸胍溶解液经分离后r-IFN-γ比活最高,SDS对r-IFN-γ的构象影响较大。

Inclusion body of recombinant human interferon gamma (r-IFN-γ) was dissolved by sodium dodecyl sulphate (SDS), urea and guanidine hydrochloride. r-IFN-γ was separa ted on gel exclusion chromatography. The effect of denaturating agents on separation results was studied by SDS-polyacrylamide gel electrophoresis, activity and high-performance hy-drophobic interaction chromatography. The difference of molecular weight between r-IFN-γ and impurity is large in urea solution of dissolving inclusion body. Specific acti vity of the r-IFN-γ separated reaches 1. O×10~8 lU/mg protein in guanidine hydrochloride as mobile phase and the agent of dissolving inclusion body and specific activity was low in SDS solution, r-IFN-γconformation change degress was large in SDS solution.