Preparation and Identification of Monoclonal Antibody Against Urease B Subunit from Helicobacter pylori

Objective:To establish the specific hybridoma cell lines that secreted stably monoclonal antibody(mAb)against urease B subunit(UreB)from Helicobacter pylori.Methods:Female BALB/c mice were immunized with purified recombinant UreB from H.pylori and mAb against UreB was prepared with hybridoma cell fusion technique.Indirect enzyme-linked immunosorbent assay(ELISA)was used to evaluate the affinity and titers of mAbs,determine their specificity,immunoglobulin subtype and identify their antigen determinants.Results:Eight stably secreting anti-UreB mAb cell lines were acquired.Cross-reactivity with Yersinia enterocolitica,Klebsiella pneumoniae and Proteus vulgaris,which also produce urease,was not detected.Their relative affinities were between 1.13×10-8～4.66×10-10.The titers of mAb in ascites were from 2×104～3.2×105.Two mAb cell lines secreted IgG1 subtype antibodies and three cell lines IgG2a antibodies.All eight antibodies belonged to three different antigen determinants.Conclusion:The specific anti-UreB mAbs of IgG1 and IgG2a subtypes were produced,which will be significant for clinical diagnosis and fundamental research in H.pylori.