摘要
在大肠杆菌Rosetta(DE3)中表达猪瘟病毒石门株NS3基因部分功能片段,以纯化的重组NS3(p80)蛋白免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经间接ELISA筛选及3次连续克隆化获得了4株可分泌抗NS3单抗的杂交瘤细胞(2G7,2F11,5D2和6F11),通过小鼠体内诱生法制备腹水,并对杂交瘤细胞和单克隆抗体的特性进行鉴定。结果表明,杂交瘤细胞染色体正常,抗体分泌稳定,单抗效价高,亲和力各不相同,Ig亚类鉴定均为IgG1。Westernblot分析证明4株单抗均能与原核表达的NS3蛋白特异性结合,6F11株与真核表达的NS3蛋白有较强的特异性反应。
Monoclonal antibodies against NS3 gene of CSFV was developed using recombinant NS3 （p80） protein expressed in E.coli Rosetta （DE3） as antigen. BALB/c mice was immunized with purified NS3 （pS0） protein and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA, subcloned and characterized. Four hybridoma cells （2G7, 2F11, 5D2 and 6F11） secreting anti-NS3 monoclonal antibodies （McAbs） were obtained. Hybridoma cells with normal chromosome were shown to stably secret NS3 specific antibodies with various affinities. All the McAbs belong to IgG1 subtype.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2008年第6期464-468,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金项目(30270988)
陕西省自然科学基金项目(2006C121)
关键词
猪瘟病毒
NS3基因
表达
蛋白纯化
单克隆抗体
classical swine fever virus （CSFV）
NS3 gene
expression
protein purification
monoclonal antibody （McAb）
