论文部分内容阅读
为获得高纯度的抗肾综合征出血热病毒单克隆抗体,建立杂交瘤细胞株7C11两步纯化的方法并进行亚类及特异性鉴定。采用Balb/c小鼠制备7C11单克隆抗体腹水,辛酸-硫酸铵法初步分离纯化,蛋白G亲和层析柱进一步分离纯化并鉴定。经初步分离纯化后,单抗IgG纯度>80%,回收率>50%;经过Protein G层析柱二次分离纯化后,单抗IgG纯度>98%,回收率>50%。亚类鉴定为IgG1,且特异性较好。成功获得高纯度的抗HFRS单克隆抗体,为抗汉坦型肾综合征出血热抗原表位的鉴定、抗原物质结构与功能的关系等后续研究奠定基础。
To obtain a high purity monoclonal antibody against hemorrhagic fever with renal syndrome (HFRS), a two-step purification method of hybridoma cell line 7C11 was established and subclassified and identified. Balb / c mice were used to prepare the ascites of 7C11 monoclonal antibody. The protein was separated and purified by caprylic acid - ammonium sulfate method. Protein G affinity chromatography was further purified and identified. After preliminary isolation and purification, monoclonal antibody IgG purity> 80%, recovery> 50%; After purification by Protein G chromatography twice, monoclonal antibody IgG purity> 98%, recovery> 50%. Subclass was identified as IgG1, and the specificity is better. The successful obtaining of high purity anti-HFRS monoclonal antibody lays a foundation for further studies on the identification of anti-Han Tan typhoid antigen epitopes and the relationship between the structure and function of antigens.