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中文摘要：
目的获得纯化人突变CD59（hmCD59）蛋白,制备其特异性抗体,对纯化产物进行鉴定。方法将含有hmCD59全长序列的重组pALTER质粒,运用脂质体介导法转染CHO细胞,采用SDS-PAGE检测hmCD59的表达情况,表达产物经ANTI-FLAG M2 Affinity Gel纯化后制备抗血清,以此抗血清鉴定纯化产物。结果hmCD59在CHO细胞获得表达,ANTI-FLAG M2 Affinity Gel纯化后可获得电泳纯的hmCD59蛋白,hmCD59与小鼠抗hmCD59抗血清具有特异性反应。结论从真核细胞获得了电泳纯并具有抗原性的hmCD59蛋白,为进一步对其进行抗体制备、功能研究及临床应用奠定了基础。
英文摘要：
Objective To obtain purified human mutant CD59 （hmCD59） protein and to identify the purified product with specific antiserum against hmCD59 protein. Methods The recombinant pALTER plasmid containing hmCD59 cDNA was transfected into CHO cells with pcDNA3 by lipofectamine. Expression of hmCD59 on CHO cells was detected by SDS-PAGE. The recombinant protein was purified by ANTI FLAG M2 Affinity Gel. The antiserum was prepared with the purified hmCD59 and used to identify the purified hmCD59. Results hmCD59 protein was purified by ANTI-FLAG M2 Affinity Gel. Furthermore, hmCD59 had specific reactivity with mouse anti hmCD59 antiserum. Conclusion The purified hmCD59 protein has been obtained successfully, which lays the foundation for preparing antibody, further functional study and clinical application of hmCD59.
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