摘要： 采用单克隆抗体制备技术，建立了分泌抗猪瘟病毒单克隆抗体的杂交瘤细胞株，液氮保存4年后复苏，细胞在HAT选择培养基中生长旺盛，分泌抗体稳定，分别命名为YNF1，YNF2，YNF3，YNF4，杂交瘤细胞染色体数介于96~115条。细胞培养上清和腹水的单抗间接ELISA效价分别为1：80~1：160和1：5 000~1：10 000。在间接ELISA和夹心ELISA检测中单抗与猪瘟病毒呈特异性反应，与正常的猪、兔肾组织提取液及其血清Ig无交叉。抗体蛋白属IgG类，亚类及轻链分别为IgG1/λ，IgG2a/κ，IgG2a/λ，IgG1/λ。结果表明，4株杂交瘤是分泌抗猪瘟病毒单克隆抗体的细胞株，可长期传代无限分泌单抗，是进一步研究猪瘟病毒和开发敏感、特异、快速诊断猪瘟试剂盒和试纸探针的免疫学特异性试剂来源。
关键词:
猪瘟病毒,
杂交瘤,
单克隆抗体,
ELISA
Abstract: The hybridoma cell lines,YNF1,YNF2,YNF3 and YNF4 secreting anti-CSFV monoclonal antibodies,were established by cell fusion technique.These cells,after preserved for four years and resuscitated,grew prosperously and secreted antibodies steadily.The chromosome number of the cells variated between 96 and 115.The ELISA titers of cell supernatant and ascites were 1:80 and 1:10000.The monoclonal antibodies are specific to CSFV and can't bind to the normal samples such as pig's kidney and liver in indirect-ELISA and sandwich-ELISA.YNF1,YNF2,YNF3,and YNF4 all belong to class IgG,and subclass IgG1/λ,IgG2a/κ,IgG2a/λ,IgG1/λ respectively.The result indicated that four hybridoma cell lines can infinitely secrete anti-CSFV monoclonal antibody that can be used as immunology reagent to develop sensitive,specific,fast diagnosis kit and reagent paper probe.
Key words:
CSFV,
Hybridoma,
Monoclonal antibody(McAb),
ELISA
中图分类号:
杨艳艳, 张改平, 肖治军, 杨继飞, 柴书军, 冯春花, 王选年. 分泌抗猪瘟病毒单克隆抗体杂交瘤细胞株的建立[J]. 华北农学报, 2006, 21(5): 60-63. doi: 10.3321/j.issn:1000-7091.2006.05.015.
YANG Yan-yan, ZHANG Gai-ping, XIAO Zhi-jun, YANG Ji-fei, CHAI Shu-jun, FENG Chun-hua, WANG Xuan-nian. The Establishment of Hybridoma Cell Lines Secreting Monoclonal Antibodies to CSFV[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2006, 21(5): 60-63. doi: 10.3321/j.issn:1000-7091.2006.05.015.