title: [Preparation of monoclonal antibody against recombinant human interferon-β1a].
abstract: OBJECTIVE
  To prepare the monoclonal antibody (mAb) against recombinant human interferon-β1a (rhIFN-β1a).
  
  
  METHODS
  BALB/c mice were immunized with purified rhIFN-β1a. The spleen lymphocyte cells from the one whose blood specific antibody titer exceeded 1:10 000 were fused with Sp2/0 cells. The fused cells went through HAT selection, serial dilution, and positive selection until stable hybridoma cell lines were obtained. The equilibrium dissociation constant (K(D)) of antibodies was calculated based on ELISA result. The cell line with the best KD value was cultured in a stirred-tank bioreactor with a fed-batch strategy to produce secreted anti-rhIFN-β1a monoclonal antibody. The mAb released into the cell culture supernatant were purified by ultrafiltration and protein-G affinity chromatography.
  
  
  RESULTS
  Thirteen hybridoma cell lines which stably produced anti-rhIFN-β1a antibody were retrieved from standard hybridoma fusion and selection procedures. One of the 13 cell lines, 1E8 with the KD value of 4.6 × 10⁻⁹ mol/L was cultured and antibody titer in the cell culture supernatant reached 1:5000. The mAb with 90% purity was recovered from the two-steps purification.
  
  
  CONCLUSIONS
  The anti-rhIFN-β1a mAb with high purity has been successfully obtained.
authors.affiliation: School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, Shenzhen 518044, China.
authors.forename: Dengyang
authors.initials: D
authors.lastname: Zhang
authors.affiliation: School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, Shenzhen 518044, China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: LA
authors.affiliation: School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, Shenzhen 518044, China.
authors.forename: Liuping
authors.initials: L
authors.lastname: Li
authors.affiliation: School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, Shenzhen 518044, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002460:Cell Line; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D000068556:Interferon beta-1a / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D000068556:Interferon beta-1a