抗delta-like-1单克隆抗体的制备与鉴定  

Preparation and identification of a monoclonal antibody against delta-like-1

作　　者：张小洪[1] 魏大鹏[1] 李天佳[1] 罗志娟[1] 

机构地区：[1]四川大学华西基础与法医学院,四川成都610041

出　　处：《华西药学杂志》2010年第6期712-714,共3页West China Journal of Pharmaceutical Sciences

摘　　要：目的制备抗delta-like-1的单克隆抗体并鉴定其特异性。方法用delta-like-1多肽-BSA蛋白复合物作为免疫原免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞(SP2/0)融合,经多次筛选及克隆化,建立可以稳定分泌抗delta-like-1单抗的杂交瘤细胞株。用ELISA、Western blot、细胞免疫组化对此抗体特性进行鉴定。结果筛选到两株能稳定分泌抗delta-like-1单抗的细胞株3H11C11A9E3和2D5D11F8F1,亚类鉴定两株均为IgG1,轻链为kappa链;ELISA法测定两株细胞腹水抗体效价分别为1:4.096×105和1:1.024×105,抗体亲和常数分别为3.98×107、3.28×107L.mol-1;Western blot显示此单抗能特异性识别神经胶质瘤细胞株U251中的天然delta-like-1蛋白;细胞免疫组化进一步显示delta-like-1分布于细胞核中。结论成功制备了抗delta-like-1单克隆抗体,可为研究delta-like-1与脑胶质瘤等神经系统疾病的关系奠定基础。OBJECTIVE To prepare and identify the monoclonal antibody(mAb) against delta-like-1.METHODS Splenocytes isolated from Balb/c mice which were immunized with a peptide of delta-like-1 coupled to bovine serum albumin(BSA) were fused with mouse myeloma cells SP2/0.After several rounds of screening and cloning,two stable strains of hybridoma were established.The specificity was evaluated with ELISA and Western blot,and the titer,immunoglobulin subtype and affinity constant of the mAbs were identified.RESULTS Two...

关 键 词：Delta-like-1 单克隆抗体 U251细胞株 酶联免疫吸附法 

分 类 号：R392[医药卫生—免疫学]