title: [Expression of human-mouse chimeric antibody against CD40 in CHO cell line and characterization of its function].
abstract: OBJECTIVE
  To investigate the stable expression of a chimeric antibody against CD40 moleculeèch-5C11éin CHO and its biological activity.
  
  
  METHODS
  Human-mouse chimeric antibody against CD40 recombinant plasmid and mock plasmid were transfected into CHO cell line through lipofectamine mediation. Human kappa chain and Fc fragment of ch-5C11 were characterized by FACS and Western blot. The concentration of ch-5C11 in cell supernatants was detected by Lowry assay and the inhibitory effect of ch-5C11 on the proliferation of Daudi cells was detected by MTT assay.
  
  
  RESULTS
  RT-PCR showed that target CHO cells integrated chimeric heavy chain and chimeric light chain gene. FACS and Western blot showed that ch-5C11 in cell supernatants maintained the binding activity and specificity to human CD40 molecule, and contained human kappa chain and Fc fragment. Cell supernatants were purified using protein G affinity chromatography. The concentration of human-mouse chimeric antibody against CD40 in cell supernatants was 0.535 mg/L. When co-cultured with B lymphoma cell line Daudi, ch-5C11 induced proliferation arrest of Daudi cells.
  
  
  CONCLUSIONS
  The human-mouse chimeric antibody against CD40 can be expressed in CHO stably and effectively, which inhibits proliferation of Daudi.
authors.affiliation: Key Laboratory of Clinical Immunology of Jiangsu Province, First Affiliated Hospital, Soochow University, Suzhou 215006, China.
authors.forename: Qiu-xia
authors.initials: QX
authors.lastname: Qu
authors.forename: Cheng
authors.initials: C
authors.lastname: Chen
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Ge
authors.forename: Yong-jing
authors.initials: YJ
authors.lastname: Chen
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Qiu
authors.forename: Xue-guang
authors.initials: XG
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D019013:CD40 Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins