抗β-淀粉样蛋白单克隆抗体杂交瘤细胞系的建立及初步鉴定  

Establishment and Primary Identification of Hybridoma Cell Line Secreting McAb against β-amyloid

作　　者：周辉[1] 徐葛林[1] 严家新[1] 盛柏杨[2] 公衍道[2] 

机构地区：[1]武汉生物制品研究所基因工程室,武汉430060 [2]清华大学生物系,北京100084

出　　处：《中国生物制品学杂志》2008年第5期410-413,共4页Chinese Journal of Biologicals

摘　　要：目的建立抗β-淀粉样蛋白（Aβ）单克隆抗体杂交瘤细胞系，并鉴定邱单抗的免疫学特性，以便用于阿尔茨海默病（AD）的防治研究。方法用戊二醛法将半抗原Aβ1-42偶联于载体蛋白BSA，形成结合抗原BSA-Aβ。用常规腹腔注射法和微量足垫注射法分别免疫BALB／c小鼠，采用杂交瘤技术建立分泌Aβ单抗的细胞株，体内诱生腹水法制备Aβ单抗。硫酸铵盐析法提纯，常规免疫学法检测鉴定Aβ单抗的免疫学特性。结果筛选出A-B7、A-D11、A-E93株高特异性杂交瘤细胞株，亚型均为Igm型，单抗的ELISA间接效价分别为2×10^-5、2×10^-5和1×10^-50杂交瘤细胞染色体数目在80-90之间，纯化后单抗腹水蛋白回收率在20％左右。结论已成功建立了分泌抗Aβ单抗的细胞系，并制备出半抗原Aβ的单抗，可用于AD的进一步研究。Objective To establish the hybridoma cell hne secreting McAb against β-amyloid and identify the produced McAb which might be used for the prevention and treatment of Alzheimer disease （AD） .Melhods Prepare conjugated antigen BSA-Aβ by coupling hapten Aβ1-42 to vector protein BSA with glutaraldehyde method. Immunize BALB/c mice with the prepared conjugated antigen by routine intraperitoneal injection and micminjection into food pad. Estabhsh the cell strains secreting McAb against Aβ by hybridoma technique and prepare the McAb by induction of ascites in vivo. The prepared McAb was purified by salting out with ammnonium sulfate and identified by routine immunological test. Resldts Three hybridoma cell strains secreting high specific McAb against Aβ, i.e. A-B7, A-D11 and A-E9, were established. All the secreted McAbs were of subtype IgM, and their titer titers were 2 x 10^-5, 2 x 10^-5 and 1 x 10^-5 respectively. All the chromosome number~ of the 3 hybridoma cell strains were 80 - 90. The recovery rate of protein in ascites after purification was about 20%. Conehmon The hybridoma cell hne secreting McAb against Aβ was successfully estabhshed, and the McAb was prepared, which might be used for the further study on AD.

关 键 词：Β-淀粉样蛋白 阿尔茨海默症 单克隆抗体 杂交瘤细胞系 

分 类 号：R392-33[医药卫生—免疫学]