Figure 2 : Expression of Pfs25, Pfs25-IMX313 and antigen-specific IgG responses in mice after immunization with viral-vectors and protein-in-adjuvant formulations.

From: Enhancing immunogenicity and transmission-blocking activity of malaria vaccines by fusing Pfs25 to IMX313 multimerization technology

Figure 2

(A) Western blot of supernatant from pENTR4-LPTOS shuttle plasmids transfected into HEK293 cells (left panel) and protein purified from P. pastoris supernatant (middle panel) for Pfs25 and Pfs25-IMX313 under reduced (R) and non-reduced (NR) conditions. Western blotting was performed using anti-Pfs25 4B7 mAb. The purified protein was also analyzed on SDS-PAGE using coomassie stain (right panel). For the western blots, 15 μL supernatant from transfected HEK293 cells and 500 ng of purified proteins from P. pastoris were loaded per lane. For Coomassie analysis, 5 μg of purified proteins were loaded. (B) Two groups (n = 5 per group) of BALB/c mice were primed with 1X108 i.u of ChAd63 (expressing Pfs25 or Pfs25-IMX313) on day 0 and boosted 8 weeks later with 1X107 pfu of MVA (expressing Pfs25 or Pfs25-IMX313) via the intramuscular (i.m.) route. Day 14 and day 70 serum samples were collected and the total IgG response against Pfs25 was measures using standardized ELISA. (C) Six groups (n = 5 per group) of BALB/c mice were immunized i.m. in parallel with 2.5 μg, 5 μg and 10 μg of soluble Pfs25 or Pfs25-IMX313 nanoparticle formulated in Alhydrogel. Mice were primed on day 0 and boosted on day 21 followed by another boost on day 42. Day 20, 41 and 62 serum were collected and analyzed by Pfs25 standardized ELISA. (D) Two groups (n = 5 per group) of BALB/c mice were immunized via the i.m. route with 2.5 μg of Pfs25 or Pfs25-IMX313 formulated in Alhydrogel. Mice were primed on day 0 and boosted on day 21. Serum was collected on days 20, 50, 80, 150, 200 and 250 and anti-Pfs25 total IgG was measured. Mean with SEM are depicted for (B–D). Individual data are shown for B and C. Mann-Whitney test was performed *p < 0.05, **p < 0.01.