棉铃虫单粒包埋核型多角体病毒粒子单克隆抗体及抗原特性分析

PREPARATION OF MONOCLONAL ANTIBODIES AGAINST HELIOTHIS ARMIGERA SINGLY EMBEDDED NUCLEAR POLYHEDROSIS VIRUS (HaSNPV) AND ANALYSIS OF VIRAL ANTIGENIC PROPERTIES

经多角体碱溶超离心和柱层析纯化的棉铃虫单粒包埋核型多角体病毒(HaSNPV)粒子作为抗原,通过细胞融合,得到3株分泌抗HaSNPV单克隆抗体的杂交瘤细胞D_2、B_8和D_7,相应抗体的效价MAD_2为10,MAB_8为50,MAD_7为1000。其中MAB_8为构象决定簇抗体,MAD_2和MAD_7为顺序决定簇抗体。用SDS-PAGE,Western-blot和ELISA方法确定MAD_2对应的抗原决定簇多肽大小为56kd和96kd,MAD_7对应56kd多肽。在不同病毒株系鉴别中发现MAD_2,MAD_8为HaSNPV特异,而MAD_7则对大袋蛾单粒包埋核型多角体病毒(CvNPV)发生交叉反应。另外用ELISA法检测了HaSNPV在体外细胞系统中的增殖动态。

Three hybridomas designated as D_2, B_8 and D_7, secreting monoclonal antibodies againstHeliothis armigera singly embeddcd nuclcar polyhedrosis virus (HaSNPV) were established by the cell fusion technique. The titers of monoclonal antibodics dcsignated as MAD_2, MAB_8 and MAD_7 were 10, 50and 1000, respectively. Of the three monoclonal antibodies, MAB_8 was the monoclonal antibody toconformational epitope, whereas MAD_2 and MAD_7 to sequential cpitopes on the enveloped virion. ByWestern-blot analysis, it was shown that MAD_2 reacted with two viral polypeptides: 56kd and 96kd, andMAD_7 with 56kd. Two different NPVs (HaSNPV and CvNPV) could be distinguished with the monoclonal antibodies prepared. The kinetics of HaSNPV multiplication in vitro was also siudied with the antibodics by ELISA.