摘要
应用大肠杆菌表达的猪瘟病毒主要保护性 E2抗原决定簇蛋白和全长 E2基因构建的 DNA疫苗免疫 BAL B/c小鼠。取免疫小鼠脾细胞与 SP2 /0骨髓瘤细胞进行融合 ,经克隆和间接 EL ISA筛选 ,获得了 A11、B2、E3、H6、D5、D86株稳定分泌抗猪瘟病毒 E2蛋白单克隆抗体 (Mc Ab)的杂交瘤细胞株。它们的腹水效价在 1∶ 80 0~ 1∶ 2 10 0 0 0之间。抗体类型鉴定结果表明 ,A11、E3、H6、D5和 D8为 Ig M类型 ,B2为 Ig G类型。随后用蛋白 A凝胶层析法和 PEG沉淀法分别纯化了 Ig G和 Ig M单抗。对纯化单抗进行的抗原识别表位研究结果初步表明 ,6株单抗可能识别 3种不同的E2抗原表位。
A panel of monoclonal antibodies(McAbs) against E2 protein of classical swine fever virus were generated by immunization of BALB/c with:a.E.coli-expressed one third part of N terminal of E2,which includes predominant neutralizing antigenic determinants,b.DNA vaccine based on entire E2 gene.All 6 hybridomas were characterized for their potency of secreting monoclonal antibody showing the antibody titers between 1∶800- 210 000 .The preliminary identification of antigenic epitopes recognized by those McAbs in ELISA by using synthetic peptides showed that these 6 McAbs could recognize three epitopes of immunogenic E2 protein.These antibodies will be used to delineate the relationship between E2 gene variation and the antigenic change of the virus.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2002年第2期121-124,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金重大项目 ( 398932 90-1)
国家"973"项目 ( G19990 119-0 3)