[Cloning and expression of Fab gene of mAb HAb18 against human hepatocellular carcinoma].
OBJECTIVE
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.


CONCLUSIONS
HAb18 Fab was prepared successfully, which lays the foundation for its further application to diagnosis and therapy of human hepatocellular carcinoma.
Xing
Jin-liang
JL
Cell Engineering Research Center, Fourth Military Medical University, Xi'an 710032, China. xinglinliang@163.com
Wang
Yong-qing
YQ
Yang
Xiang-min
XM
Song
Fei
F
Chen
Zhi-nan
ZN
D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000276:immunology; D008113:Liver Neoplasms / Q000276:immunology*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments