摘要
为了解猪链球菌2型(SS2)菌株毒力相关因子的致病机理,建立高效的免疫学检测方法,将SS2胞外因子(EF)抗原性强的区域通过基因克隆、原核表达,经亲和层析纯化的EF蛋白免疫BALB/c小鼠,采用B淋巴细胞杂交瘤技术将免疫细鼠脾细胞与SP2/0骨髓瘤细胞融合,建立了4株分泌抗EF蛋白单克隆抗体(McAb)的杂交瘤细胞系2G1、5H7、3H4和1F2。间接ELISA测定杂交瘤细胞培养上清的效价为1∶128～1∶512,诱生腹水的抗体效价为1∶104～1∶105。4株McAb腹水的Western blot鉴定表明McAb能特异性的识别EF。以2G1单抗腹水与和EF多克隆抗血清建立的夹心ELISA可特异地检测EF。
To investigate the pathogenesis mechanisms of Streptococcus suis type 2 （SS2） toxicity related factors and develop highly effective diagnostic methods, the dominant antigenic region of extracellular factor protein （EF） gene fragment was cloned into prokaryotic expression plasmid. The recombinant protein was purified by affinity chromatography and used to immuize BALB/c mice. Four hybridomas 2G1, 5H7, 3H4 and 1F2 secreting monoclonal antibodies （McAb） to EF were established by fusion of mouse myeloma cells SP2/0 with spleen cells from the immunized BALB/c mice. The antibody titer was measured with indirect ELISA which ranged from 1：128-1：512 in culture supematants and 1：104-1：105 in ascitic fluids. The four McAb recognized EF specifically by western blot. A sandwich ELISA for the detection of EF was developed utilizing 2G1 McAb and EF polyclonal antibody.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2008年第7期562-565,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家“十五”重大传染病科技攻关计划(No.2003BA712A03-05)
江苏省自然科学基金(No.BK2006014)
关键词
猪链球菌2型
胞外因子
单克隆抗体
鉴定
Streptococcus suis serotype 2
extracellular factor
McAb
identification
