title: [Preparation, identification and application of monoclonal antibody against urease subuint B of Helicobacter pylori].
abstract: OBJECTIVE
  To prepare and characterize the monoclonal antibody (mAb) against Helicobacter pylori (Hp) and establish a competitive ELISA used for detection of Hp antibodies in the sera of Hp-infected patients.
  
  
  METHODS
  BALB/c mice were immunized with inactivated Hp to generate Hp mAb using the hybridoma technology. Hp mixed proteins including cytotoxin-associated gene A (CagA), vacuolating cytotoxin A (VacA) and urease, as well as inactivated Hp were applied to screen positive hybridoma. Selected Hp mAb was analyzed and characterized with ELISA and Western blotting, and then labeled with horseradish peroxidase (HRP) for establishing a competitive ELISA to detect Hp antibodies in the sera of Hp-infected patients.
  
  
  RESULTS
  One Hp mAb named as C3 was selected after screening large amount of hybridoma, and the C3 Hp mAb was special for IgG2a subtype with the affinity titer of 1×10(7). Western blotting, ELISA and mass spectrum analysis indicated that the C3 Hp mAb could recognize Hp urease subunit B specifically. Using the C3 Hp mAb, we developed a competitive ELISA which could be used to detect Hp antibodies in the sera of Hp-infected patients.
  
  
  CONCLUSIONS
  We successfully obtained one mAb that could specifically recognize Hp urease subunit B and developed a competitive ELISA using the Hp mAb.
authors.affiliation: Hybridoma and Antibody Center, School of Life Science, Xiamen 361005, China.
authors.forename: Huandi
authors.initials: H
authors.lastname: Ding
authors.affiliation: Department of Clinical Laboratory, First Affiliated Hospital, Xiamen University, Xiamen 361005, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Han
authors.affiliation: Hybridoma and Antibody Center, School of Life Science, Xiamen 361005, China.
authors.forename: Angen
authors.initials: A
authors.lastname: Huang
authors.affiliation: Hybridoma and Antibody Center, School of Life Science, Xiamen 361005, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Meng
authors.affiliation: Hybridoma and Antibody Center, School of Life Science, Xiamen 361005, China.
authors.forename: Huaxin
authors.initials: H
authors.lastname: Hu
authors.affiliation: Hybridoma and Antibody Center, School of Life Science, Xiamen 361005, China.
authors.forename: Xiaotong
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016480:Helicobacter pylori / Q000201:enzymology / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D021122:Protein Subunits; D014510:Urease / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D021122:Protein Subunits; D014510:Urease