摘要
用恶性疟原虫MSP1 31基因片段的重组质粒DNA直接免疫BALB/c小鼠 ,诱导产生体液 ,免疫后取脾细胞与SP2 /0小鼠骨髓瘤细胞在PEG1 450作用下进行融合 ,获得了 2株能分泌抗恶性疟原虫MSP1 31单克隆抗体的小鼠杂交瘤细胞株 9H9和 8A2。用酶联免疫吸附试验检测 ,小鼠腹水抗体滴度最高为 1∶1 0 0 0 0。经免疫球蛋白类型和亚类鉴定 ,2株杂交瘤细胞株均为IgG1 。蛋白免疫印迹试验表明 ,此单克隆抗体与MSP1 31蛋白抗原有特异免疫反应 ,证明通过质粒DNA直接免疫小鼠可制备特异性单克隆抗体。
BALB/c mice were immunized by injection of recombinant plasmid DNA carrying MSP1\|31 fragment of \%Plasmodium falciparum.\% After antiserum was induced, McAbs were prepared by fusing SP2/0 myeloma cells with spleen cells. Specific McAbs were detected in hybridoma culture supernatant and mouse ascites by ELISA, positive hybridomas were cloned by technique of limiting dilutions. Two McAbs, 9H9 and 8A2, against the malaria protein were produced and characterized. The titers of the two McAbs in acites were 1∶ 10000 and 1∶ 2500 respectively. The Ig subclass of these McAbs was IgG\-1. Western blot test showed that the McAb specifically reacted with MSP1-31 antigen. These results proved that monoclonal antibodies could be produced by use of immunization of mice with plasmid DNA or DNA vaccine.
出处
《微生物学报》
CAS
CSCD
北大核心
2000年第4期389-393,共5页
Acta Microbiologica Sinica
基金
上海复星高科技 (集团 )有限公司资助
关键词
严重疟原虫
DNA疫苗
单克隆抗体
制备
载体DNA
Plasmodium falciparum, DNA vaccine, Monoclonal antibodies, Hybridoma, Genetic immunization
