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抗腺病毒载体单克隆抗体的制备及其初步鉴定
引用本文:张春燕,李方和,龚劲松,陈妍,李时君,黄永国,张波. 抗腺病毒载体单克隆抗体的制备及其初步鉴定[J]. 华中科技大学学报(医学版), 2008, 37(2): 255-258. DOI: 10.3870/j.issn.1672-0741.2008.02.031
作者姓名:张春燕  李方和  龚劲松  陈妍  李时君  黄永国  张波
作者单位:1. 华中科技大学同济医学院附属同济医院实验医学研究中心,武汉,430030
2. 武汉生物制品研究所,武汉,430060
摘    要:
目的 研制抗腺病毒载体(Adv)表面蛋白的单克隆抗体(McAb).方法 将Adv以Al(OH)3佐剂化,常规免疫BALB/c小鼠.采用细胞融合技术制备抗Adv McAb;采用ELISA、免疫细胞化学及Western blot法对McAb进行筛选与鉴定.结果 共获得6株可稳定分泌特异性McAb的细胞(A4H11、A8C7、F1H5、G1D2、G4E3、H2G8),其染色体数目为102~106条,所分泌McAb均属IgG1亚类,持续3个月适应培养仍能保持其稳定的抗体分泌能力.腹水抗体的ELISA效价在1∶106~1∶108之间;Western blot结果显示6株McAb均与来自腺病毒载体及野生3、5、7型腺病毒抗原提取物中分子量约为114 kD的多肽反应,据此推测它们均是抗ADV-TK六邻体蛋白的McAb.结论 成功制备出6株抗腺病毒载体六邻体单克隆抗体,为腺病毒载体应用于肿瘤基因治疗的临床前研究提供物质基础.

关 键 词:基因治疗  腺病毒载体  单克隆抗体  毒载体  单克隆抗体  Adenovirus Vector  Monoclonal Antibodies  Identification  物质基础  临床前研究  基因治疗  肿瘤  应用  蛋白  六邻体  反应  多肽  分子量  提取物  抗原  病毒  显示  腹水
修稿时间:2007-06-15

Preparation and Identification of Monoclonal Antibodies against Adenovirus Vector
Zhang Chunyan,Li Fanghe,Gong Jingsong et al Research Center of Experimental Medicine,Tongji Hospital,Tongji Medical College,Hauzhong University of Science , Technilogy,Wuhan. Preparation and Identification of Monoclonal Antibodies against Adenovirus Vector[J]. Journal of Huazhong University of Science and Technology(Health Sciences), 2008, 37(2): 255-258. DOI: 10.3870/j.issn.1672-0741.2008.02.031
Authors:Zhang Chunyan  Li Fanghe  Gong Jingsong et al Research Center of Experimental Medicine  Tongji Hospital  Tongji Medical College  Hauzhong University of Science    Technilogy  Wuhan
Affiliation:Zhang Chunyan,Li Fanghe,Gong Jingsong et al Research Center of Experimental Medicine,Tongji Hospital,Tongji Medical College,Hauzhong University of Science , Technilogy,Wuhan 430030
Abstract:
Objective To prepare and identify monoclonal antibodies(McAb) against the capsid proteins of adenovirus vector.Methods BALB/c mice were immunized with a mixture of the purified adenovirus vector(Adv) and Al(OH)3.McAbs were produced using cell fusion technique in a conventional way.The sensitivity and specificity of McAb were identified by indirect enzyme linked immunosorbent assay(ELISA),immunocytochemical staining and Western blot.Results Six strains of hybridoma cells(A4H11,A8C7,F1H5,G1D2,G4E3 and H2G8) t...
Keywords:gene therapy  adenovirus vector  monoclonal antibodies  
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