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Fig. 6 | Microbial Cell Factories

Fig. 6

From: Immunogenicity of Leishmania-derived hepatitis B small surface antigen particles exposing highly conserved E2 epitope of hepatitis C virus

Fig. 6

Analysis of the humoral response induced by Leishmania-derived particles in BALB/c mice. a Analysis of the antibody endpoint titers of the pooled mouse antisera specific to the recombinant particles. ELISA plates were coated with 412–425_sHBsAg (right) or sHBsAg (left) particles. b Analysis of the interaction of immune sera with yeast-derived HBsAg proteins. ELISA plates were coated with 5 µg/mL of purified HBsAg protein from P. pastoris (yHBsAg) (right) or commercially available vaccine against HBV (Engerix-B) (left). c Analysis of the antibody response to the 412–425 synthetic peptide. ELISA plates were coated with 20 µg of 412–425 peptide. AP33 mAb was used to estimate concentration of Abs specific to the 412–425 region in serum. Dilution factor of the pooled 412–425_sHBsAg sera and concentration of the AP33 antibody are shown on x-axis (bottom and top, respectively). The 412–425_sHBsAg sera values are represented with bars, the AP33 values are marked with solid line. Mean A450 values and standard deviations are shown on the y-axis. The background from the negative control serum in each dilution was subtracted from the obtained results (a, b, c). For each ELISA assay, the mean from three independent experiments performed in duplicate is shown. Asterisks indicate statistical significance (*P < 0.05, paired two-tailed t-test) (a, b). The solid horizontal line (a, b) indicates the cutoff value (three times the mean background value). d Analysis of cross-reactivity of the 412–425_sHBsAg sera to the E1E2 complex from different HCV genotypes. The figure represents western blotting in reducing conditions with 412–425_sHBsAg sera diluted 1:500. As an antigen, extracts of HEK293 cells transfected with plasmids expressing E1E2 glycoproteins from different HCV genotypes were used. Non-transfected HEK293 cell lysate was used as negative control (NC)

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