从包涵体中纯化重组人幽门螺杆菌尿素酶B亚单位

目的 建立一种有效的方法，从包涵体中纯化重组人幽门螺杆菌尿素酶B亚单位（rUreB）。方法重组大肠杆菌发酵后，表达的rUreB包涵体经洗涤、变性、复性，采用Q Sepharose Performance阴离子交换层析和Pheny1 Sepharose High Performance疏水层析分离纯化。使用 SDS－PAGE和HPLC检测纯度，选用 ELISA和 Western－bloning对纯化蛋白的生物学活性进行鉴定。结果 rUreB包涵体经洗涤和溶解后，rUreB的纯度＞70％。包涵体溶解液经阴离子交换层析和疏水层析后纯度超过95％。rUreB纯品具有良好的生物学活性。结论 建立了从包涵体中获得高纯度的rUreB的工艺，为进一步的研究打下了基础。

Purification of Recombinant Human Helicobacter pylori Urease B from Inclusion Body

Abstract] Objective To establish an effective method for purifying recombinant human Helicobacter pylori urease B (rUreB) from inclusion body. Methods The inclusion bodies expressed in E. coli were washed, denatured and renatured, then the rUreB in them were purified by Q Sepharose High Performance ion exchange and Phenyl Sepharose Fast Flow chromatography. The purity of rUreB was detected by SDS - PAGE and HPLC, and the biological activity of it by ELISA and Western blot. Results The purity of rUreB in inclusion bodies after being washed and dissolved was above 70% . However, after being purified by the two - step chromatography procedure, the purity reached above 95%. The purified rUreB showed good biological activity. Conclusion An effective method for purifying rUreB from inclusion body is established. It lays a foundation of further study in animals.