目的：探索原核表达重组人骨形成蛋白 -2(rhBMP-2)的纯化和复性方法。方法：对携带有pBV220-hBMP-2的 工程 菌进行发酵，收集菌体，超声破碎菌体后制备包涵体。包涵体经进一步超声裂解后过凝胶色 谱和离子交换色谱，对获得的高纯度蛋白进行不同条件下的复性。rhBMP-2产物作用于培养 的人牙乳头细胞，检测ALP活性。结果：产物经过凝胶色谱后纯度达 到90%，复性后再经离子交换和凝胶色谱纯度为85%以上，并且提高了生物学活性。结论：rhBMP-2产物经过离子交换色谱和凝胶色谱获得较高纯度的重组蛋白； 3mol/L 尿素和0.75～1.25mol/L的NaCl能显著提高产物生物学活性。%AIM: To study methods for purificat ion and regeneration of E.coli generated rhBMP-2. METHODS: TOP10 strain carrying pBV220-rhBMP-2 recombinant construct was fermented and lysised to prepare inclusion bodies which was then further lysised by ultrosonic followe d by Gel chromatography and ion-chromatographyprocessures to generate high puri ty protein product. different regenerative conditions were investigated to obtai n rhBMP-2 product of higher biological activity measured by ALP activity analysi s. RESULTS: The purity of primitive rhBMP-2 product increa sed by 90% after Gel chromatography and the regenerated protein increased its pu rity by 85% after ion-exchange chromatography. CONCLUSIONS: Gel chromatography and ion-exchange Chromatogaphy help obtain rhBMP product of high purity; 3mol/L urea and 0.75～1.25mol/L NaCl can regenerate rhBMP-2 to a higher level of biological activity.
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