title: [Characterization and preliminary application of six monoclonal antibodies against recombinant signal protein 14-3-3 of Schistosoma japonicum].
abstract: OBJECTIVE
  To identify the properties of six monoclonal antibodies (McAbs) against recombinant signal protein 14-3-3 of Schistosoma japonicum, and investigate their value in the diagnosis of schistosomiasis.
  
  
  METHODS
  The subclasses, titers, affinity-constants, detection limits and specificities of six McAbs were identified by ELISA and Western blotting. Dot Enzyme-linked Immunosorbent Assay (Dot-ELISA) for detecting the 14-3-3 protein in the sera of rabbits infected with Schistosoma japonicum was established, then it was used to observe the dynamics of 14-3-3 protein in sera of rabbits infected with Schistosoma japonicum before and after treatment. The diagnostic value of Dot-ELISA was investigated through detecting a group of sera samples of rabbits infected with Schistosoma japonicum.
  
  
  RESULTS
  The types of six McAbs against recombinant signal protein 14-3-3 of Schistosoma japonicum of 3F1, 3F7, 5C6, 5D1, 5G9 and 1G6 were all IgG, their subclasses were IgG1, IgG2a, IgG2b, IgG1, IgG1 and IgG1, their antibody-titers in ascites were 1:6.4 x 10(5), 1:8.0 x 10(5) , 1:6.4 x 10(5), 1:3.2 x 10(5), 1:4.8 x 10(5) and 1:2.0 x 10(4), their affinity-constants were 8.82 x 10(8), 4.93 x 10(8), 1.56 x 10(8), 5.12 x 10(8), 1.41 x 10(8) moL/L and 2.30 x 10(7) mol/L, their detection limits in dot-ELISA were 1, 10, 100, 10, 10, 100 ng, respectively. All the six McAbs recognized the recombinant signal protein 14-3-3 of Schistosoma japonicum and the natural signal protein 14-3-3 in SEA, ESA and AWA, and did not react with the protein of E. coli and Clonorchis sinensis in Western blotting. 3F1 and 5D1 were selected to establish the Dot- ELISA for detecting the sera of rabbits infected with Schistosoma japonicum before and after treatment at different time points, it was found that the concentration of 14-3-3 protein in sera of rabbits was increased gradually with time and decreased gradually after the infected rabbits treated with praziquantel. Forty-two sera samples of rabbits infected with Schistosoma japonicum were detected by this Dot-ELISA, 41 samples were positive, the sensitivity was 97.6%, and all of the ten healthy rabbits' sera were negative, the specificity was 100%.
  
  
  CONCLUSIONS
  All the six McAbs could recognize natural signal protein 14-3-3. It has been proved preliminarily that the Dot-ELISA based on 3F1 and 5D1 is valuable for the diagnosis of active infection of Schistosoma japonicum and accessing the chemotherapeutic effect of schistosomiasis.
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: Chuan-Xin
authors.initials: CX
authors.lastname: Yu
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Song
authors.forename: Xun-Ren
authors.initials: XR
authors.lastname: Yin
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Yong-Liang
authors.initials: YL
authors.lastname: Xu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
mesh_terms: D048948:14-3-3 Proteins / Q000097:blood / Q000276:immunology*; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas; D007106:Immune Sera; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D012549:Schistosoma japonicum / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000175:diagnosis* / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D048948:14-3-3 Proteins; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D007106:Immune Sera; D007074:Immunoglobulin G; D011994:Recombinant Proteins