摘要
用提纯的黄瓜绿斑驳花叶病毒(CGMMV)免疫的BALB/c鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,经细胞筛选与克隆,获得4株能稳定传代并分泌抗CGMMV单克隆抗体(MAb)的杂交瘤细胞,并分别制备其单抗腹水。4株单克隆腹水抗体间接ELISA效价在10-6～10-7之间,4株单抗的抗体类型及亚类均为IgG1,kappa链。特异性检测表明4株单抗仅与CGMMV的17.5 ku的外壳蛋白有特异性反应。利用最灵敏的11E5单抗为核心建立了检测CGMMV抗原包被间接ELISA方法(ACP-ELISA),该方法对病叶的检测灵敏度达到1∶10 240(g/mL)倍稀释,对提纯病毒的检测灵敏度达到0.01 ng。
After selected and cloned, four hybridoma cell lines secreting monoclonal antibodies（MAbs） against Cucumber green mottle mosaic virus（CGMMV） were produced by fusing mouse myeloma cells（SP2/0） with spleen cells from BALB/c immunized by CGMMV particles. The ELISA titres of ascitic fluids of four MAbs ranged from 10^-6-10%-7. Isotypes and subclasses of this four MAbs all belonged to IgG1, Klight chain. All four MAbs could specifically react with coat protein （17.5 ku） of CGMMV. Based the most sensitive MAb, an indirect antigen-coated plate （ACP）-ELISA for CGMMV detection was set up. The sensitivity analysis indicated the ACP-ELISA based the MAb 11E5 could successfully detect virus in plant leaves sap at 1：10240（g/mL）dilution or 0.01 ng purified CGMMV.
出处
《热带作物学报》
CSCD
2010年第7期1162-1166,共5页
Chinese Journal of Tropical Crops
基金
浙江省自然科学基金重点项目(No.Z3090039)
