title: [Construction of expression vector for human-mouse chimeric antibody and expression of an antibody against human HER2].
abstract: OBJECTIVE
  To construct universal eukaryotic expression vector for expressing human-mouse chimeric antibody and have application in expression of PCR-obtained mouse variable region gene fragments in the form of human-mouse chimeric antibody, for the purpose of clinical therapy.
  
  
  METHODS
  By using human Tac antigen signal peptide and kappa chain and gamma 1 heavy chain gene fragments of human Ig, we constructed a versatile expression vector for human-mouse chimeric antibody and used it to transfect cultured 293T cells. The expressed product was detected using RT-PCR, FACS and ELISA.
  
  
  RESULTS
  An expression vector for PCR-derived V genes of mouse Ig as human-mouse chimeric antibody was constructed. The V(L) and V(H) genes encoding mouse anti-human HER2 antibody were amplified with primers designed for this expression system, and an expression vector for chimeric antibody against human HER2 was constructed. By transfection of 293T cells, we demonstrated that a chimeric antibody was produced and secreted to the culture medium.
  
  
  CONCLUSIONS
  This expression system is useful for antibody engineering for it provides a simple way to express chimeric antibody from V genes of mouse antibody obtained by PCR.
authors.affiliation: Institute of Modern Separation Sciences, Northwest University, Xi'an 710069, China.
authors.forename: Li-li
authors.initials: LL
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Han
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002460:Cell Line; D004622:Embryo, Mammalian; D004847:Epithelial Cells / Q000378:metabolism; D018734:Genes, erbB-2; D005822:Genetic Vectors; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics; D007668:Kidney / Q000166:cytology; D051379:Mice; D021382:Protein Sorting Signals / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D015375:Receptors, Interleukin-2 / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007142:Immunoglobulin gamma-Chains; D021382:Protein Sorting Signals; D018719:Erb-b2 Receptor Tyrosine Kinases; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins