猪囊尾蚴特异性单克隆抗体的制备及其识别抗原成分的鉴定

Preparation and immunological characteristics of monoclonal antibodies to Cysticercus cellulosae

通过SDS-PAGE方法对猪囊尾蚴囊液抗原成分进行了分离鉴定,并分别分离纯化了其中的16 000和10 000特异性蛋白质成分;将16 000和10 000纯化蛋白质成分分别做成佛氏佐剂苗,分组免疫BALB/c小鼠,超免后,无菌取脾脏制备脾细胞,分别与NS0骨髓瘤细胞进行融合,经阳性筛选和特异性鉴定获得2株分泌猪囊尾蚴特异性单抗的杂交瘤细胞,命名为TSCF-1611H12B8和TSCF-1012G5B5。免疫学鉴定结果表明,单抗TSCF-1611H12B8和TSCF-1012G5B5与猪细颈囊尾蚴、旋毛虫、住肉孢子虫和蛔虫抗原间不存在交叉反应;单抗TSCF-1611H12B8识别囊液中16 000和10 000蛋白质抗原条带,而单抗TSCF-1012G5B5识别囊液中的10 000蛋白质条带。

In order to establish Hybridoma cell lines secreting antibodies specific to Cysticercus cellulosae, the cystic fluid of Cysticercus cellulosae was separated by SDS-PAGE. Two specific protein contents, which were 16 000 and 10 000 respectively, were obtained, and then applied to two groups of BALB/c mice after being mixed with Freund＇s adjuvant. Spleen cells were collected from the immunized mice and then fused with NS0 myeloma cell line. Two strains of hybridoma cells secreting antibodies specific to Cysticercus cellulosae, were acquired and named TSCF-1611H12B8 and TSCF-1012G5B5. Monoclonal antibodies TSCF-1611H12B8 and TSCF-1Q2G5B5 showed no cross-reactivity with any antigens from rrichinella spiralis, Cysticercus tenuicollis, Sarcocystis rniescheriana or Ascaris suurn. Epitope mapping showed that monoclonal antibody TSCF-1611H12B8 identified 16 000 and 10 000 proteins while only 10 000 protein was identified by TSCF-1012G5B5.