title: [Preparation of monoclonal antibodies against the adhesion protein 33 of Trichomonas vaginalis].
abstract: OBJECTIVE
  To prepare and characterize the monoclonal antibodies (McAbs) against recombinant adhesion protein 33 (AP33) of Trichomonas vaginalis.
  
  
  METHODS
  The purified recombinant fusion protein AP33 was used as antigen to immunize BALB/c mice. Sp2/0 myeloma cells were fused with the splenocytes from immunized BALB/c mice. After ELISA screening and 4 times of limited dilution, 5 positive hybridoma cell lines were obtained, and the biological properties of the McAbs were identified by Western blotting. Indirect immunofluorescent antibody test (IFAT) was performed and the inhibition effect of McAbs on the cytoadherence of Trichomonas vaginalis to HeLa cell was assayed.
  
  
  RESULTS
  Western blotting demonstrated that 5 McAbs, designated as 4A2, 4F11, 4F8, 4E7 and 4H11, specifically combined with the recombinant AP33 of T. vaginalis. The McAbs were IgG1 isotypes. Four of them (4F11, 4F8, 4E7 and 4H11) showed parasite recognition by IFAT. Parasite cytoadherence to a monolayer of HeLa cells was inhibited in vitro with a inhibition rate of 50.08%, 65.03%, 50.70% and 49.08% by the 4 McAbs under a concentration of 200, 200, 400 and 200 microg/ml, respectively.
  
  
  CONCLUSIONS
  The prepared McAbs against the recombinant AP33 show a protective inhibition on cytoadherence of Trichomonas vaginalis in vitro.
authors.affiliation: Wenzhou Medical College, Wenzhou 325035, China.
authors.forename: Hui-Cong
authors.initials: HC
authors.lastname: Huang
authors.forename: Shi-Fang
authors.initials: SF
authors.lastname: Yu
authors.forename: Ming
authors.initials: M
authors.lastname: Cai
authors.forename: Feng
authors.initials: F
authors.lastname: Tan
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Zheng
authors.forename: Chang-Wang
authors.initials: CW
authors.lastname: Pan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D015800:Protozoan Proteins / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014246:Trichomonas vaginalis / Q000187:drug effects / Q000276:immunology*
chemical_list: C113260:AP33 protein, Trichomonas vaginalis; D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins