title: [Cloning and expression of Fab gene of mAb HAb18 against human hepatocellular carcinoma].
abstract: 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.
authors.affiliation: Cell Engineering Research Center, Fourth Military Medical University, Xi'an 710032, China. xinglinliang@163.com
authors.forename: Jin-liang
authors.initials: JL
authors.lastname: Xing
authors.forename: Yong-qing
authors.initials: YQ
authors.lastname: Wang
authors.forename: Xiang-min
authors.initials: XM
authors.lastname: Yang
authors.forename: Fei
authors.initials: F
authors.lastname: Song
authors.forename: Zhi-nan
authors.initials: ZN
authors.lastname: Chen
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments