摘要
将纯化的安氏隐孢子虫卵囊超声波破碎,反复冻融后免疫BALB/C小鼠,取其脾细胞与SP2/0细胞融合,经3~5次有限稀释克隆化,筛选出4株持续分泌抗体的杂交瘤细胞株.通过ELISA,IFA和EITB测其反应性.结果表明,4株杂交瘤细胞培养上清效价为1∶100-1∶12 800,诱生腹水效价为1∶6 400~1∶102 400;4株单抗均特异性识别安氏隐孢子虫卵囊壁抗原;免疫印迹表明,4株单抗分别与38.0~97.2 kD的主要抗原蛋白条带起反应.
BALB/C mice were immunized with purified oocyst antigens of C.andersoni which had been freezen and thawed for several times and treated by ultrasonic wave.Spleen cells of the mice were selected to fuse with SP2/0 cells.The positive wells were selected by indirect ELISA,and after three to five generations of cloning four hybridoma cell clones were obtained.The characterization of McAbs was studied by ELISA,IFA and EITB.Results indicated that the supernant titers ranged from 1∶100 to 1∶12 800 and ascites titers from 1∶6 400 to 1∶102 400.Examined with IFA,all monoclonal ascites identified oocyst wall antigen.Western blot poofile shows that the four McAbs could be recognized respectively by 38.0～97.2 kD polypeptide antigen bands of C.andersoni oocysts.
出处
《河南农业大学学报》
CAS
CSCD
北大核心
2010年第1期65-68,共4页
Journal of Henan Agricultural University
基金
卫生部公益性行业科研项目(30871863)
