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用核酸免疫技术制备抗丙肝病毒C区单克隆抗体
引用本文:李益民,周旭.用核酸免疫技术制备抗丙肝病毒C区单克隆抗体[J].微生物学免疫学进展,1998,26(1):64-65.
作者姓名:李益民  周旭
作者单位:卫生部兰州生物制品研究所
摘    要:用丙肝病毒C+E1区真核表达质粒pcDNA-HCV/C+E1按400ug/只剂量免疫BALB/C小鼠,14周后同一剂量再加强免疫一次。加强免疫后2周,在50%PEG1450介导下将脾细胞与SP2/0小鼠骨髓瘤细胞(51)融合。实验结果融合率达54.3%(313/576)。阳性率为5.4%(17”313)。克隆化后得到6株稳定分泌抗丙肝病毒C区单克隆抗体杂交瘤细胞株。这6株杂交瘤均产生IgM抗体,接种BALB/C小鼠后产生腹水的效价为164-1320(ELISA)。

关 键 词:丙肝病毒  核酸免疫  单克隆抗体  杂交瘤

Preparation of Monoclonal Antibodies against Hepatitis C/C by Immunizing Mice with DNA
Abstract:The BALB/C mouse was immunized with 400 ug hepatitis C virus/C+E1 eukaryotic expression plasmid-pcDNA HCV/C+E1.After 14 weeks,the mouse was boosted with the same amount of pcNDA-HCV/C+E1 plasmid.On the 14th day after last immunization,the fusion of the spleen cells and SP2/0 myloma cells (5∶1) was carred out.The fusion produced 313 wells of hybridoma.Of these growing wells,5.4% (17/313) contained antibody against gene-recombinant HCV/C antigen. After clonning,6 strains of hybridoma were obtained. All of these produced IgM to HCV/C. The titers of the antibodies in ascites were 1∶64 to 1∶320 (ELISA).
Keywords:hepatitis C  DNA immunization  monoclonal antibody  Hybridoma
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