摘要
以纯化复性的原核表达的番鸭呼肠孤病毒(MDRV)σB蛋白为抗原免疫BALB/c雌性小鼠,按常规方法制备单克隆抗体,通过间接ELISA、Western-blot和免疫组织化学试验进行筛选及鉴定。结果显示,获得4株能够稳定分泌抗σB蛋白单克隆抗体的杂交瘤细胞株(A3F、B2B、C3E和D2G),其亚型分别属于IgG2bI、gG2bI、gM和IgM。采用小鼠体内诱生法制备的腹水均能与MDRV的σB蛋白发生特异性反应,表明,制备的单克隆抗体能够识别天然构象的σB蛋白。
σB protein of Muscovy duck reovirus（MDRV） was expressed in Escherichia coli BL21（DE3） harboring pET30-σB plasmid. Female BALB/c mice were immunized by using the purified σB protein as immunogen to screen monoclonal antibodies（mAbs） against the σB protein by using indirect enzyme linked immunosorbent assay, Western-blot, and immunohistoehemistry assay. The results showed that four strains of hybridoma cell lines stably secreting mAbs against σB, designated as A3F, B2B,C3E, and D2G respectively,were obtained, and they belonged to subtype IgG2b, IgG2b, IgM, or IgM respectively. The four mAhsin the ascites of mice inoculated with the four hybridoma cell lines could react specifically with the σB protein expressed in MDRV-infected duck embryo fibroblasts,indicating that the mAbs could recognize the σB protein in natural conformation.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2009年第9期786-789,共4页
Chinese Veterinary Science
基金
国家重点基础研究发展计划(973)项目(2005CB522905)
关键词
番鸭呼肠孤病毒
σB蛋白
单抗
Muscovy duck reovirus
σB protein
monoclonal antibody
