单克隆抗体的制备、纯化和保存方法对其特异性的影响

来源 :国外医学.预防.诊断.治疗用生物制品分册 | 被引量 : 0次 | 上传用户:cliff800
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将流感病毒NT68株所产生的血凝素作为抗原,通过杂交瘤技术获得38个亚克隆细胞株.其中各有10株分别分泌IgG_1、IgG_(2a)、IgG_(2b)、8株分泌IgM.杂交瘤细胞经组织培养或注入BALB/c小鼠腹腔制备单克隆抗体McAb.用同种病毒作血凝抑制(HI)试验测定抗体效价,用一组非同种病毒株作HI试验测定McAb的特异性和交叉反应性.McAb经以下物理处理:(1)温度:将腹水作1∶500~1∶2000系列稀释后,分别置62℃、10分钟;56℃、30分钟;37℃、1小时;20℃、16小时.并以 Thirty-eight subclone cell lines were obtained by hybridoma technique using hemagglutinin derived from influenza virus NT68 as antigen, of which 10 had secreted IgG1, IgG2a, IgG2b and 8 secreted IgM, respectively. The hybridoma cells were cultured in vitro or injected into the peritoneal cavity of BALB / c mice to prepare monoclonal antibody McAb.The antibody titer was determined by using the same virus as the hemagglutination inhibition (HI) test, and the McAb (1) Temperature: The ascites was serially diluted 1: 500 to 1: 2000 and then set at 62 ° C for 10 minutes, 56 ° C for 30 minutes, 37 ° C, 1 hour; 20 ℃, 16 hours
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