title: [Preparation of monoclonal antibodies against multiple antigens].
abstract: OBJECTIVE
  To prepare monoclonal antibodies (mAbs) against multiple antigens by single cell fusion.
  
  
  METHODS
  BALB/c mice were immunized with the multiple antigens, namely alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), HBsAgiHBcAg and HBeAg, and hybridomas were employed using PEG as the fusing agent. The hybridoma cells were respectively screened with AFP, CEA, HBsAg, HBcAg and HBeAg by enzyme-linked immunosorbent assay and limited dilution. The mAbs were purified by protein G affinity chromatography, its subtype was identified, the affinity constants (K(a)) were determined and the specificity was analyzed by Western blotting.
  
  
  RESULTS
  Twenty hybridoma cell lines were obtained by single cell fusion, including 5 cell lines against AFP, 6 against CEA, 3 against HBsAg, 4 against HBcAg, and 2 against HBeAg. The subtypes of some hybridoma cell lines positive for the mAbs were identified as the immunoglobulin G1 (IgG1), with K(a) ranging from 1x10(9) M(-1) to x10(11) M(-1). Western blot analysis showed that all the mAbs strongly and specifically bound to their respective antigens.
  
  
  CONCLUSIONS
  The mAbs against multiple antigens have been obtained by single cell-fusion, which increases the production of mAbs and reduces the time of preparation.
authors.affiliation: Institute of Tropical Medicine, Southern Medical University, Guangzhou 510515, China. dannypen@163.com
authors.forename: Dan-dan
authors.initials: DD
authors.lastname: Peng
authors.forename: Yun-shan
authors.initials: YS
authors.lastname: Ning
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yan-hua
authors.initials: YH
authors.lastname: Hong
authors.forename: Yun-dan
authors.initials: YD
authors.lastname: Wang
authors.forename: Fen-fang
authors.initials: FF
authors.lastname: Wu
authors.forename: Ming
authors.initials: M
authors.lastname: Li
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D002459:Cell Fusion / Q000379:methods; D005260:Female; D006511:Hepatitis B Antigens / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006513:Hepatitis B e Antigens / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D000509:alpha-Fetoproteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D000509:alpha-Fetoproteins