重组人血管生长素包涵体复性条件研究

目的摸索重组人血管生长素(rhANG)复性影响条件,以提高rhANG的复性效率。方法超声波破碎菌体,变性萃取rhANG包涵体。分析rhANG包涵体起始浓度、氧化型、还原型谷胱甘肽比例、盐酸胍和添加精氨酸对复性的影响。结果在0.1 mol/L Tris-HCl pH 7~8,还原型、氧化型谷胱甘肽比率为5,盐酸胍为0.2~0.5 mol/L,起始蛋白质浓度0.1~0.2 mg/mL时,rhANG蛋白复性率为60%;0.5 mol/LL-精氨酸有助于复性。结论初步优化rhANG包涵体复性参数,为rhANG的放大制备创造了条件。

Study on renaturation parameters of inclusion bodies of recombinant human angiogenin

Purpose To investigate the renaturation parameters of inclusion bodies of recombinant human angiogenin(rhANG) for improving the refolding efficiency of rhANG.Methods Cells were disrupted by sonication,and inclusion bodies of rhANG were extracted in denature reagents.The effect of GSH/GSSG ratio,pH,concentration of protein,guanidine hydrochloride and L-arginine on the refolding yield of rhANG was studied.Results When the refolding buffer included 0.1 mol/L Tris-HCl pH 7-8,(GSH/GSSG)=5,0.2-0.5 mol/L guanidine hydrochloride and 0.1-0.2 mg/mL rhANG,the refolding efficiency of rhANG was 60%.Adding 0.5 mol/L L-arginine can enhance the refolding yield.Conclusion The renaturation parameters of inclusion bodies of rhANG were preliminarily optimized which provide a basis for rhANG scale-up production.