摘要
目的:克隆抗人肝癌单抗(mAb)HAb18的Fab基因,并在大肠杆菌中表达。方法:采用RT-PCR法,利用设计的引物扩增mAb Fd和K链全长基因,并分别克隆到T载体中进行序列分析。然后,亚克隆到原核表达载体pComb3中,转化感受态大肠杆菌后以IPTG诱导表达。采用ELISA和免疫荧光法检测表达产物的特异性。结果:克隆了mAb HAb18的Fah基因,并在大肠杆菌中获得表达。竞争性ELISA和免疫荧光检测证实,表达产物具有与相应抗原特异结合的活性。结论: 成功地构建了抗人肝癌小分子Fab抗体,为其进一步在肝癌诊断与治疗中的应用奠定了基础。
AIM: To clone Fab gene of mAb HAb18 against human hep-atocellular carcinoma and express it in E. coli. METHODS: The cDNAs of K chain and Fd of mAb HAb18 were amplified by RT-PCR and cloned into prokaryotic expression vector pComb3 which was transfected into competent E. coli to express Fab by IPTG induction. The specificity of the expressed Fab was tested by ELISA and immunofluorescence staining. RESULTS: The Fab gene of mAb HAb18 was successfully amplified and expressed in ? coli. The results of competitive ELISA and immunofluorescence staining showed that the expressed Fab had specific antigen-binding activity. CONCLUSION: HAb18 Fab was prepared successfully, which lays a foundation for its further application to diagnosis and therapy of human hepatocellular carcinoma.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2003年第6期570-573,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家高技术研究发展计划(863)重点课题(No.2001AA215101)
关键词
肝癌
单克隆抗体
FAB
表达
hepatocellular carcinoma
monoclonal antibody
Fab
expression
