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. 2019 May 28;10:1185. doi: 10.3389/fimmu.2019.01185

Figure 5.

Figure 5

Neutralizing antibodies against urease and adherence inhibition assay. (A) Purification of anti-FVpE IgG antibodies. Anti-FVpE antibodies were purified by Sepharose 4B to which the purified FVpE was attached through stable covalent bonds. After purification, anti-FVpE antibodies were detected by SDS-PAGE. (B) H. pylori urease neutralization test. H. pylori urease was pre-incubated with the IgG antibodies (60 μg/well) against FVpE, CTB-UE, or CTB. After addition of phenol red indicator, the color development was measured at 550 nm at 30 min intervals over a period of 3 h. The data are expressed as percentage inhibition. (C) Specific antibodies against the UreB321−339 epitope peptides of the urease active site. ELISA plates were coated with 1 μg/well of synthetic UreB321−339 peptides. ***p < 0.001, ns, not significant. (D) Adherence inhibition assay. GES-1 cells were cultured with H. pylori pretreated with various concentrations of IgG from mice immunized with FVpE, CTB-UE, or CTB. After stained with Giemsa, the number of adherent H. pylori was counted by an oil-immersion microscope. These results were verified in triplicate assays. *p < 0.05 compared with mouse anti-CTB antibody.