title: [Construction of eukaryotic vector and expression of chimeric antibody against rh-bFGF].
abstract: OBJECTIVE
  To express a human-mouse chimeric antibody against rh-bFGF antigen in eukaryotic cells.
  
  
  METHODS
  The VL and VH genes were amplified and cloned from the hybridoma 1F11 secreting anti-rh-bFGF mouse monoclonal antibody (mAb) and the CL and CH genes were amplified and cloned from the plasmid pMDHC and pMDLC. They were inserted into eukaryotic expression vectors after sequencing. The recombinant plasmids were then transformed into CHO-dhfr- cells for expression. The humanization and antigen specificity of expressed products were identified by indirect ELISA. The relative molecular mass (Mr) of expressed products was determined by SDS-PAGE.
  
  
  RESULTS
  The cloned antibody genes were identified to be functional by sequencing. The chimeric antibody could be detected in the culture supernatant of transformed CHO cells. The humanization and specificity of the chimeric antibody to rh-bFGF were confirmed by ELISA.
  
  
  CONCLUSIONS
  The mouse-human chimeric antibody against rh-bFGF has been expressed successfully in eukaryotic cells, which lays the foundation for its further clinical research.
authors.affiliation: Lab. of Molecular Immunology and Antibody Engineering, College of Life Sciences and Technology, Jinan University, Guangzhou 510632, China. txjj@jnu.edu.cn
authors.forename: Jun-jian
authors.initials: JJ
authors.lastname: Xiang
authors.forename: Hong-qing
authors.initials: HQ
authors.lastname: Li
authors.forename: Ning
authors.initials: N
authors.lastname: Deng
authors.forename: Hong-yu
authors.initials: HY
authors.lastname: Yang
authors.forename: Yi-gang
authors.initials: YG
authors.lastname: Tong
authors.forename: Jian-min
authors.initials: JM
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D050438:Genes, Immunoglobulin Heavy Chain; D050439:Genes, Immunoglobulin Light Chain; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D016222:Fibroblast Growth Factor 2