【摘要】 目的　获得抗华支睾吸虫循环抗原Fab段抗体。方法　将华支睾吸虫代谢抗原包被在固相ELISA板中 ,采用“吸附 -洗脱 -扩增”的方法 ,对已构建的抗华支睾吸虫噬菌体抗体库进行富集。从富集后的抗体库中筛选抗循环抗原抗体克隆 ,并交叉筛选鉴定其特异性。结果　从 2 0 6个克隆中筛选到阳性克隆 2 3个 ,再分别用肺吸虫、肝片虫、姜片虫和日本血吸虫脱脂全虫粗抗原进一步交叉筛选 ,最终获得特异性抗Cs -MAg阳性克隆 3个 (B0 5 ,C10和E38)。对其中 2株 (E38和B0 5 )序列分析表明它们的κ轻链V基因属于人VκⅠ (O2 )亚群 ,J基因属于Jκ1;γ链V基因属于人VHⅠ - 6 9亚群 ,D基因来自D3- 10 ,J基因则来自JH3。结论　用成虫代谢抗原直接包板 ,可以从抗体库中筛选到阳性克隆 ,构建的噬菌体抗体库是成功有效的更多还原

【Abstract】 Aim To obtain Fab antibodies against Clonorchis sinensis CAg Methods Cs MAg was used in panning from phage library against Clonorchis sinensis The MAg binding activity and specificity of phage antibodies were assayed by ELISA Results 206 random clones were tested for the presence of anti Cs MAg Fabs,and 23 clones showed positive The specificity of these 23 clones was confirmed by binding them to other four trematode antigens(Paragonimus westermani,Fasciola hepatica,Fasciolopsis buski,Schistosoma japonicum) There were 3 clones(B05,C10,E38) bound to Cs MAg but not to any of the other four trematode antigens Comparison of the Fd gene and light chain gene of E38 and B05 with GenBank and KABAT database showed that the heavy chains belonged to human IgG1 subclass,and their variable regions were derived from rearranged germ line gene of V HⅠ 69,D3 10 and J H3 while the light chains were a member of VκⅠ family,the germ line genes were V κⅠ(O2) and J κ1 Conclusions Phage Fab antibodies against Clonorchis sinensis CAg are selected from antibody library It also suggests that the phage antibody library is constructed successfully更多还原