作者简介: 张佳锴,男,硕士研究生,主要从事肿瘤免疫学研究,
Tel: 0592-2180587，E-mail: baroce@ 163.com; 庄国洪，通信作者，E-mail:
[email protected]。[中文摘要]目的构建、表达、纯化抗人死亡受体5(DR5)单链抗体,并检测其诱导肿瘤细胞凋亡的活性。方法采用RT-PCR获取鼠源性抗人DR5单克隆抗体重链和轻链可变区基因序列,以一柔性连接肽连接二者,转入表达载体,以大肠杆菌表达融合蛋白,亲和色谱纯化后用MTT实验和凋亡检测试剂盒检测其凋亡诱导活性。结果获得的序列经比对为抗体重链和轻链可变区基因,表达纯化后的重组蛋白具有接近完整抗体的肿瘤细胞凋亡诱导活性。结论抗人DR5 scFv可作为诱导肿瘤细胞凋亡的候选药物,为肿瘤免疫学研究提供材料。[英文摘要]Purpose To construct，express，purify anti-human DR5 scFv，and to assay its activity of
apoptosis induction for tumor cell lines． Methods Variable region sequences of heavy chain and light chain to murine anti-human DR5 monoclonal antibody were acquired by RT-PCR， then they were linked through a flexible linker peptide and was cloned into the expression vector． After being expressed by E．coli and purified using affinity chromatography， the recombinant protein was employed to MTT assay as well as apoptosis assay kit in order to detect its apoptosis-inducing activity． Results The sequences we've got were established as the variable region genes of antibody's heavy and light chain，while the recombinant proteins expressed and purified possessing the apoptosis-inducing activity come near to complete antibody．Conclusion Anti-human DR5 scFv can supply material for tumor immunology research as drug candidate inducing tumor cell apoptosis．福建省自然科学基金资助项目(C0710046
