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长叶车前花叶病毒单克隆抗体的制备及在病毒分型中的应用

PRODUCTION OF MONOCLONAL ANTIBODIES TO RIBGRASS MOSAIC VIRUS AND APPLICATION IN SEROTYPING

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【作者】 张永平李洸李德葆

【Author】 Zhang Yongping Li Debao Li Guang Zhejiang Agricultural University, Hangzhou Zhejiang Academy of Medicine,Hangzhou

【机构】 浙江农业大学生物技术研究中心浙江医学研究院免疫研究室浙江农业大学生物技术研究中心 杭州杭州杭州

【摘要】 应用细胞杂交技术,将长叶车前花叶病毒杭州分离株(RMVha)、烟草花叶病毒普通株(TMVc)和北京番茄株(TMVbe-t)免疫的Balb/c小鼠脾细胞与Sp2/0-Ag14小鼠骨髓瘤细胞融合,经筛选测定,成功地获得了5株稳定分泌特异性单克隆抗体的杂交瘤细胞株,其中1株属于IgG2b,其余4株均属于IgG?,5株细胞株均制备了腹水抗体,ELISA效价最高达256000,琼脂双扩散效价达128,根据与15个不同RMV和TMV毒株的反应特性,可将5株单克隆抗体分为A、B、C三组,分别针对a、b、c三个不同的抗原决定簇。根据三组单克隆抗体反应性的差异,15个毒株可分为8个血清型,本文讨论了所获得的5株杂交瘤细胞在植物病毒的诊断、病毒病原的鉴定及病毒分型方面的应用价值。

【Abstract】 Five stable hybridoma cell lines ( Rm1, Rm2, Rm3, Rm4 and Rm5 ) which secrete monoclonal antibodies of ribgrass mosaic virus ( RMY ) and/or tobacco mosaic virus ( TMV ) were produced by fusing spleen cells of mice immunized with either RMVha or a mixture of RMVha, TMVc anb TMVbe-t to mouse myeloma cell line Sp2/0-Ag14. It was found that the five monoclonal antibodies represented two isotypes of murine antibodies, IgG2b and IgG8. High-titered ascitic fluid was produced with all of the hybridomas.Three antigenic determinants (epitopes), six serotypes were proposed by the authors after testing these monoclonal antibodies with 15 isolates of RMV and TMV. Serotype Ⅰ(TMVbe-t and TMVna-L2)has at least the determinants a, b and c; Serotype Ⅱ (TMVc, TMVgu-t and TMVbe-1)the dete- rminants b and c, SerotypesI (TMVbe-1.2 ) ,Ⅳ ( RMVha, RMVsh, TMV-Ⅱ and TMVna-B2)and V (TMVnc-0, TMVsh-0, TMVxi-0 and TMV-Ⅰ ) have the determinants c, a and b, respectively; Serotype Ⅵ ( TMVsh-1.2 ) , however, does not have any of these determinants, probably has determinants other than the a, b and c. The potential value of the monoclonal antibodies in virus research is discussed.

  • 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,1987年02期
  • 【被引频次】4
  • 【下载频次】26
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