Fig. 2

Characterization of the particles expressed in the L. tarentolae system. a Immunofluorescence of recombinant L. tarentolae cells transfected with plasmids expressing 412–425_sHBsAg and sHBsAg. Cells transfected with empty pLEXSY_I-blecherry3 plasmid were used as negative control (NC). The staining was carried out using AP33 mAbs (blue) and anti-HBsAg Abs (green), scale bar = 5 µm. b Western blot analysis of the Leishmania-derived particles in reducing conditions. sHBsAg and 412–425_sHBsAg were treated with PNGase F and detected using the anti-HBsAg specific antibody. G represents the glycosylated and DG deglycosylated form of protein. c Recognition of particles with anti-HBsAg and AP33 Abs in the ELISA tests. ELISA plates were coated with serial dilutions of recombinant L. tarentolae cell lysates containing sHBsAg and 412–425_sHBsAg particles. The dilution factor is depicted on x-axis. For each ELISA assay, the mean from three independent experiments performed in duplicate is shown. The mean A₄₅₀ values and standard deviations are shown on the y-axis. The background from the L. tarentolae wild-type cell lysate in each dilution was subtracted from the obtained results