重组骨形态发生蛋白-7的复性及纯化

目的探讨重组骨形态发生蛋白-7（rhBMP-7）的复性及纯化获得高纯度rhBMP-7二聚体的方法。方法表达rhBMP-7的工程菌经发酵、裂菌、提取包涵体后,4℃下依次在尿素浓度为4、2、1 mol/L的磷酸盐缓冲液中透析使蛋白复性,然后再通过层析色谱纯化去除杂蛋白。结果纯化后rhBMP-7以单体和二聚体形式存在,纯度达95%以上,其中rhBMP-7二聚体含量可达60%以上。结论初步建立了先复性然后在低脲浓度下纯化的方法,能够获得含量较高的rhBMP-7二聚体。

Renaturation and purification of recombinant human bone morphogenetic protein-7

Objective To investigate the process of obtaining high concentration rhBMP-7 dimeride by renaturation and purification. Methods The inclusion body was extracted through cracking the expression ceils after fermentation of the Eseheriehia coli strain expressing rhBMP-7. The rhBMP-7 was dialyzed in 4℃ in phosphate buffer saline which contained urea, and the concentration of urea was 4 mol/L, 2 mol/L, 1 mol/L in turn, then the renaturated rhBMP-7 was purified by SP-FF chromatography. Results The rhBMP-7 monomer and dimeride were both existed after purification, the purity of rhBMP-7 was about 95%, and the content of rhBMP-7 dimeride was above 60%. Conclusion The process of purification in low urea concentration after renaturation was initially established. It was able to ohtained high concentration rhBMP-7 di meride.