Aim: To clone Fab gene of mAb HAb18 against human hepatocellular carcinoma and express it in E.coli.
Methods: The cDNAs of kappa 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 E.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 the foundation for its further application to diagnosis and therapy of human hepatocellular carcinoma.