摘要
用含1%甲醛的磷酸盐缓冲液对相思子毒素-a(abrin-a)减毒处理制备类毒素,以类毒素为免疫抗原分4次免疫3只BALB/c小鼠。采用聚乙二醇(PEG)法,取免疫脾细胞与小鼠骨髓瘤细胞(SP2/0)融合,筛选分泌抗abrin-a的单克隆抗体(McAb)杂交瘤细胞株。将阳性细胞株接种BALB/c小鼠制备McAb腹水,根据亚类鉴定结果,分别采用蛋白A或蛋白G亲和层析柱纯化腹水,间接酶联免疫吸附试验(ELISA)检测McAb特异性,Western-blot法分析McAb抗原识别位点。结果获得1G5、3C2和4G1共3株McAb杂交瘤细胞株,均为抗abrin-a的特异性McAb,与相思子凝集素、蓖麻毒素(ricin)和蓖麻凝集素均无交叉反应,其中4G1为特异性结合abrin-a A链的McAb,1G5和3C2两株为特异性结合abrin-a B链的McAb;制备的McAb可用于abrin-a含量的检测。
Three BALB/c mice were immunized four times with formalin toxiods prepared by attenuating the abrin-a in PBS containing 1% formaldehyde.Using polyethylene glycol(PEG) method,the immunized splenocytes were isolated and fused with SP2/0 cells.Screened hybirdoma cells,which secrete antibodies against abrin-a,were injected into the abdominal cavity of mice.Using Protein A affinity chromatography or protein G affinity chromatography,the antibodies were purified from ascites after the subtype were determined.Indi...
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第7期836-839,共4页
Chinese Journal of Veterinary Science
基金
吉林大学农学部博士科研启动基金资助项目(2006)
