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作 者:李永亮[1] 田美娜[1] 卢曾军[1] 杨苏珍[2] 刘在新[1]
机构地区:[1]中国农业科学院兰州兽医研究所、家畜疫病病原生物学国家重点实验室、农业部畜禽病毒学重点实验室、国家口蹄疫参考实验室,兰州730046 [2]河南省农业科学院动物免疫学重点实验室,郑州450002
出 处:《中国生物工程杂志》2008年第11期63-66,共4页China Biotechnology
基 金:国家“973”计划资助项目(2005CB523201)
摘 要:目的:用活体骨髓瘤细胞SP2/0做融合提高融合率制备单克隆抗体,并与常规方法比较效果。方法:将SP2/0细胞打到8周龄的SPF级BALB/c小鼠皮下,待实体瘤生长到直径达2~3cm时无菌解剖取实体瘤,分离出骨髓瘤细胞进行融合。同时用培养基培养SP2/0细胞进行融合做比较,分两组进行。比较两种方法的融合率以及两种方法制备出来的单克隆抗体的相对亲和力。结果:做了6次融合,实体瘤融合组融合率为70.4%,常规法融合组44.6%,两种方法制备单抗的相对亲和力均达到1∶100000以上。结论:利用活体实体瘤细胞进行融合能明显提高细胞融合率。Objective:Prepare monoclonal antibody (MAb) using SP2/0 cells isolated from tumor in live mouse, and compare the efficiency of cell fusion with conventional method. Methods:The SP2/0 cells were hypodermic injected in the back of the SPF level BALB/c mouse which were 8 week old and re-prepared by isolation from the tumor when it reached 2 -3cm diameter. The fusion efficiency of SP2/0 cells from live tumor and from cell culture was compared by fusing with spleen cells. The relative affinity of MAbs made by two methods were also compared. Results:Six fusions have been made. The fusion rate is 70.4% using SP2/0 ceils from live tumor, while that is 44.6% using SP2/0 cells from cell culture. The relative affinity of MAbs made by two method were all above 1:100 000. Conclusion: The fusion efficiency can be significantly improved by fusion spleen cells with SP2/0 cells from live tumor.
关 键 词:单克隆抗体(MAb)SP2/0细胞 活体骨髓瘤细胞 融合
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