Antigenic reactivity of recombinant structural proteins.

<p>Lysates of cells transfected with pTT5-P12A3C, co-transfected with pTT5-VP0, pTT5-VP3 and pTT5-VP1, mock transfected cells or purified iFMDV were analyzed by ELISA with four MAbs (1–5,2–4,3–2,3–3) directed against conformational epitopes of FMDV. Error bars are standard deviation from three independent experiments.</p

Schematic representation of FMDV genomic fragments cloned into the pTT5-vector.

<p>Schematic representation of FMDV genomic fragments cloned into the pTT5-vector.</p

Analysis of humoral response of BALB/c mice immunized with recombinant proteins.

<p>The results were obtained from the average of seven sera in each group. Mean ± SD sample is shown.</p

Western blot analysis of recombinant structural proteins.

<p>Lysates of cells transfected with pTT5-P12A3C, co-transfected with pTT5-VP0, pTT5-VP3 and pTT5-VP1 or mock transfected were analyzed by SDS-PAGE and Western blot using anti-FMDV guinea pig serum (1:500). iFMDV was used as positive control. The expected migration positions of VP0, VP3 and VP1 are indicated by arrows.</p

Recombinant empty capsids recognition by sera from vaccinated bovines.

<p>Percentage of positivity of bovine sera analyzed by ELISA for the recognition of recombinant empty capsids and mock transfected cell lysates. Percentage of positivity was calculated as (A₄₉₂ from recombinant protein or mock lysate/A₄₉₂ from iFMDV) × 100. Each pair of bars represents the sera from an individual animal.</p

Assembly of recombinant structural proteins into subviral particles.

<p>Lysates of cells transfected with pTT5-P12A3C, mock transfected cells or iFMDV were loaded onto 45-15% sucrose gradients. Fractions were collected and analyzed by solid phase ELISA. Fractions of iFMDV were measured at 260 nm for RNA detection. The known positions of the FMDV virions (146S), empty capsids (75S), pentamers (12S) and protomers (5S) are indicated.</p

Western blot analysis of recombinant proteins produced after transfection with pTT5-P12A3C or with pTT5-P12A.

<p>Lysates of cells transfected with pTT5-P12A3C, pTT5-P12A or mock transfected were analyzed by SDS-PAGE and Western blot using anti-FMDV guinea pig serum (1:500). iFMDV was used as positive control. The expected migration positions of P12A, VP0, VP3 and VP1 are indicated by arrows.</p