摘要
本文采用杂交瘤技术得到能分泌伪狂犬病病毒(PrV,Pseudorabies Virus)特异抗体的杂交瘤细胞。细胞融合率为98％,分泌特异抗体的杂交瘤细胞达65％,得到不同类和亚类的杂交瘤细胞系,克隆出的杂交瘤细胞系染色体数在82～109条之间。微量血清中和试验表明这些细胞系分泌的抗体为非中和性抗体。利用单抗IgG1与荧光素结合,制备出伪狂犬病病毒特异的荧光抗体,建立了伪狂犬病免疫荧光诊断法。同时,利用放射自显影和SDS-PAGE对病毒结构多肽进行了分析,表明伪狂犬病病毒主要有13种多肽成分,能被高免血清识别的有5种。
Hybridoma cells secreting specific antibodies to Pseudorabies virus ( PrV ) were obtained by using hybridoma techniques as described in the paper. The rate of cell fusion was up to 98%. The positive cell lines secreting specific antibodies against PrV were 65.2%. Hybridoma cell lines secreting different classes and classes of antibodies ( IgG, IgM ) had been prepared. The number of chromosomes in subcloned cell lines was between 89 and 109. Microneutralizaiton test showed that the antibodies secreted by these cell lines were not able to neutralize virus. FA conjugates were prepared by McAb 1B11 or lB4 labelled with isothiocyanate and the method of immunoiluor-escence was established for diagnosis of animal Pseudorabies. We also analyzed the structure of polypeptides of PrV by autoradiography and SDS-PAGE and found that there were 13 major polypeptides of PrV, 5 of which could be recognized by hyper immunoserum of PrV.
基金
陕西省科委
杨陵农业科技开发基金88J(2)002资助
关键词
伪狂犬病病毒
单克隆抗体
Pseudorabies virus
antibodies,monoclonal
fluorescence antibody (FA )
SDS-PAGE
