title: [Construction and expression of anti-human CD3 chimeric antibody gene and preliminary study of expressed product].
abstract: OBJECTIVE
  To construct and express anti-human CD3 chimeric antibody.
  
  
  METHODS
  The genes of variable regions of the light chain (V(L)) and heavy chain (V(H)) were cloned respectively into the expression vectors (V(L) Express, V(H) Express), and co-transfected into COS-7 cells. Expression level of the chimeric antibody in culture supernatant was detected by ELISA. The antibody was purified through protein A affinity chromatography and identified by Western blot. Binding activity of the chimeric antibody to the antigen was determined by FACS. Biological activity of the chimeric antibody was determined by mixed T-lymphocyte culture test.
  
  
  RESULTS
  The expression vectors were constructed and the anti-human CD3 chimeric antibody was expressed and purified successfully. Western blot showed that the purified antibody was human-mouse chimeric antibody. FACS result showed that the chimeric antibody had antigen-binding activity. Mixed T-lymphocyte culture test showed that the chimeric antibody could suppress proliferation of T lymphocytes.
  
  
  CONCLUSIONS
  The anti-human CD3 chimeric antibody has been constructed and expressed successfully, which lays the foundation for its further study.
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Lu
authors.forename: Chun-Ning
authors.initials: CN
authors.lastname: Lai
authors.forename: Ming
authors.initials: M
authors.lastname: Yu
authors.forename: Ning
authors.initials: N
authors.lastname: Guo
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Bei-Fen
authors.initials: BF
authors.lastname: Shen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D017252:CD3 Complex / Q000276:immunology*; D019556:COS Cells; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D011993:Recombinant Fusion Proteins