离子交换色谱法对大肠杆菌表达的人重组骨形态发生蛋白-7的纯化

重组大肠杆菌高量表达重组人骨形态发生蛋白-7(rhBMP-7)，每升培养液约得到湿菌体3g，其中目的蛋白约占菌体总蛋白量的40％。裂解离心，用低浓度变性剂洗涤初步纯化包涵体，上清中无目的蛋白损失；将包涵体溶解于高浓度变性剂溶液中，目的蛋白纯度提高到60％；然后在不同条件下用离子交换色谱法对变性状态下的蛋白质进行纯化，绝大部分杂蛋白被除去，目的蛋白纯度达95％以上；改变条件，可以减少rhBMP-7损失；用Western blot对目的蛋白进行特异性鉴定。

Purification of recombinant human bone morphogenetic protein-7 on ion exchange chromatography

E.coli was genetically engineered to produce recombinant human bone morphogenetic protein-7 (rhBMP-7) in a non-active aggregated form using a temperature-inducible expression system. 3 g cell wet weight per liter of culture broth was obtained. After washing with low concentration denaturant, the protein was purified up to 60%. Further purification through ion exchange chromatography could yield rhBMP-7 with purification exceeding 95%. Western blot was used to comfirm the protein.