Fig 4.

Recombinant protein-specific IgG1 and IgG2a antibodies determined by ELISA. Microtiter plates were coated as follows: (A) 100 ng well⁻¹ of the recombinant proteins administered individually or a pooled protein sample containing 33.3 ng well⁻¹ of each protein included in the cocktail vaccines (C1 and C2), totaling 100 ng, and (B) 100 ng well⁻¹ of each protein from the cocktail vaccines. Recombinant proteins were incubated with sera from immunized mice (diluted 1:50). Goat anti-mouse IgG1 and IgG2a antibodies supplied by the IsoQuick kit for mouse monoclonal isotyping (Sigma) were used as secondary antibodies. The data represent the mean OD₄₉₂ (n = 8) of the mouse sera collected at 105 DPI minus the mean OD₄₉₂ of the mouse sera collected at 0 DPI. All reactions were performed in triplicate. Different letters indicate significant differences (P < 0.01) among treatments. Statistical analyses were performed in GraphPad Prism 4 software using the Tukey test.