抗血管紧张素Ⅰ杂交瘤细胞株制备方法的建立  

The Establishment of a Method for Preparing Anti-AngiotensinⅠHybridoma Cell Line

作　　者：白云鹏 曾玲 徐彬瑗 申海燕 郝育杰 周洋 BAI Yunpeng;ZENG Ling;XU Binyuan;SHEN Haiyan;HAO Yujie;ZHOU Yang(Beijing North Institute of Biological Technology, Beijing100076, China)

机构地区：[1]北京北方生物技术研究所有限公司研发中心,北京100076

出　　处：《标记免疫分析与临床》2021年第6期1055-1060,共6页Labeled Immunoassays and Clinical Medicine

摘　　要：目的制备抗血管紧张素Ⅰ(angiotensin I,AⅠ)的杂交瘤细胞株,比较不同连接载体产出杂交瘤细胞的效果。方法通过对载体分别为牛血清白蛋白(BSA)和卵清蛋白(OVA)的AⅠ-BSA和AⅠ-OVA免疫原进行小鼠免疫,将骨髓瘤细胞与免疫合格的小鼠进行脾细胞融合,筛选出阳性杂交瘤细胞株,经过克隆、扩大培养、腹水诱生等工序,生产出AⅠ单克隆抗体,测定并比较不同连接载体产出杂交瘤细胞抗体的性能,筛选具有应用价值的抗体。结果本方法共产出9株AⅠ杂交瘤细胞株,其中AⅠ-OVA组产出4株,AⅠ-BSA组产出5株,进行各细胞抗体的功能检验,测定亚型、亲合常数,与AⅡ、1~7氨基酸肽段、5~8氨基酸肽段以及1~9氨基酸肽段均无交叉反应,优选出AⅠ-BSA组的D11C12单抗建立检测方法进行性能评价,空白限为0.04ng/mL,低值样本的分析内精密度为4.5%,高值样本的分析精密度为3.5%,与对照试剂盒进行68例样本检测结果相关性对比,r为0.9728。结论成功建立抗血管紧张素Ⅰ杂交瘤细胞株制备方法,可以满足AⅠ单克隆抗体的使用要求。Objective To establish a method for the preparation of AngiotensinogenⅠhybridoma cell line which was produced by different junction vectors,and the results were compared.Methods We used immunogens of AⅠ-BSA and AⅠ-OVA to immunize the mice.Myeloma cells were fused with spleen cells from immunocompetent mice,further through the process,in which the positive Hybridoma cell lines being screened,cloning,expanding the culture,and antibody preparation.We acquired the production of monoclonal antibody at last,and the performances of hybridoma cell antibodies produced by different ligating vectors were measured and compared,also the antibodies with application value were screened.Results There were 9 AⅠhybridoma cell lines,4 in AⅠ-OVA group and 5 in AⅠ-BSA group.Preliminary functional test of antibody showed results as following:subtype and affinity constant were determined.There was no cross reaction with AⅡ,1-7,5-8 and 1-9 amino acids,while the blank limit was 0.04ng/mL,the internal precision of low value samples was 4.5%,and the precision of high value samples was 3.5%.The correlation between the detection results of D11C12 McAb and the control kit was 0.9728.Conclusion The preparation method of AⅠhybridoma cell line can meet the requirement of AⅠmonoclonal antibody.

关 键 词：杂交瘤细胞株 融合 克隆 分析精密度 

分 类 号：R392-33[医药卫生—免疫学]