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目的:制备CD20的单克隆抗体(mAb),并探讨对治疗B细胞淋巴瘤的作用机制。方法:构建重组表达质粒pGEX-4T-1-CD20和pET-32a-CD20。以融合蛋白CD20-GST为免疫源免疫BALB/c雌性小鼠制备(mAb)。结果:CD20-GST和CD20-His蛋白在大肠杆菌中可高效表达,经纯化后可获得高纯度的CD20蛋白。结论:成功制备高纯度CD20蛋白并以此为抗原制备了1株能够稳定分泌抗人CD20的鼠单mAb的杂交瘤细胞株BD11F4。临床试验结果表明,抗CD20单克隆抗体既可单独应用,亦可作为放射性同位素或细胞毒制剂的载体,其疗效显著,且使用安全,副反应小。许多研究报道已阐明了抗CD20单克隆抗体治疗B细胞淋巴瘤的几种可能的效应机制。目前,人们正致力于探索一些新的治疗策略以期提高这种疗法的特异性,减少其非特异性的毒副作用。
OBJECTIVE: To prepare a monoclonal antibody (mAb) for CD20 and to explore its mechanism of action in the treatment of B-cell lymphoma. Methods: Recombinant plasmids pGEX-4T-1-CD20 and pET-32a-CD20 were constructed. BALB / c female mice were immunized with the fusion protein CD20-GST as immunogen (mAb). RESULTS: CD20-GST and CD20-His proteins were highly expressed in E. coli. After purification, highly purified CD20 protein was obtained. CONCLUSION: A high-purity CD20 protein was successfully prepared and used as an antigen to prepare a hybridoma cell line BD11F4 capable of stably secreting anti-human CD20 murine mAb. Clinical trial results show that anti-CD20 monoclonal antibody can be used alone, but also as a radioactive isotope or cytotoxic agent carrier, the effect is significant, and safe to use, with fewer side effects. A number of studies have demonstrated several possible mechanisms by which anti-CD20 monoclonal antibodies can treat B cell lymphomas. At present, people are working to explore some new treatment strategies in order to improve the specificity of this therapy and reduce its non-specific side effects.