金属螯合亲和层析法对重组幽门螺杆菌尿素酶B亚单位的纯化

背景：幽门螺杆菌（H.pylori)疫苗是近年研究的热点。H.pylori尿素酶B亚单位（UreB)是理想的候选抗原。目的：获得纯化的重组H.pylori UreB(rUreB)，为进一步的H.pylori疫苗制备打下基础。方法：采用金属螯合亲和层析法（MCAC）在变性条件下纯化rUreB,SDS-聚丙烯酰胺凝胶电泳（PAGE）和免疫印迹法鉴定纯化产物的分子量、纯度和抗原性。结果：纯化产物的分子量与预计分子量相符，具有良好的特异性UreB免疫原性，且一步即可达到90％以上的纯度。结论：MCAC是一种简单、高效的rUreB纯化方法。

Purification of Recombinant Helicobacter pylori Urease B Subunit Using Metal Chelated Af-finity Chromatography

Background: Helicobacter pylori (H. pylori) has been commonly studied in recent years. Urease B subunit (UreB) is an ideal antigen candidate. Aims: To obtain purified recombinant H. pylori UreB (rUreB) as the basis of H. pylori vaccine production. Methods: rUreB was purified by metal chelated affinity chromatography (MCAC) under denaturing conditions. SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblotting were used to evaluate the molecular weight, purity and antigenicity of the purified rUreB. Results: The purified rUreB, whose molecular weight was consistent with the predicted one, could recognize the specific anti-UreB antibody, and the purity attained in one step was greater than 90%. Conclusions: MCAC is a simple and rapid method for purification of rUreB.