title: [Epitope identification of monoclonal antibody 5C6 against 14-3-3 protein of Schistosoma japonicum].
abstract: OBJECTIVE
  To identify the epitope of monoclonal antibody (McAb) 5C6 against 14-3-3 protein of Schistosoma japonicum by phage display peptide library.
  
  
  METHODS
  The phage display 12-peptide library was screened with purified McAb 5C6 against 14-3-3 protein of S. japonicum three rounds of bio-panning "adsorption-elution-amplification" to enrich the specific binding phages. The single phage clones selected randomly were amplified, their genomic DNA were extracted and sequenced. The immune response characterization of phages with the same or high homologous foreign inserted DNA sequence was identified by ELISA and Western blotting for further defining the epitope recognized by McAb 5C6.
  
  
  RESULTS
  A total of 33 single phage clones were selected and sequenced. Among them, 25 shared the same foreign inserted DNA sequence of 5'-CCACCTAGTAGCAGACCGATTCTCAGTCGAAGGAAA-3', encoding a deduced peptide PPSSRPILSRRK. This peptide was not homologue to Sj14-3-3 protein or any other known native protein in the world. The results of Western blotting showed that this peptide could be recognized by the sera of patients with schistosomiasis, but not by those of healthy persons.
  
  
  CONCLUSIONS
  The mimic antigen epitope of McAb 5C6 against 14-3-3 protein of S. japonicum, which is a conformational epitope, has been defined successfully in this study.
authors.affiliation: Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Wuxi 214064, China.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Qian
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Song
authors.forename: Chuan-Xin
authors.initials: CX
authors.lastname: Yu
authors.forename: Xu-Ren
authors.initials: XR
authors.lastname: Yin
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Jin
authors.forename: Xue-Dan
authors.initials: XD
authors.lastname: Ke
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D001483:Base Sequence; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D019151:Peptide Library; D012549:Schistosoma japonicum / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D048948:14-3-3 Proteins; D000911:Antibodies, Monoclonal; D000939:Epitopes; D019151:Peptide Library