MOESM5 of Generation of Plasmodium falciparum parasite-inhibitory antibodies by immunization with recombinantly-expressed CyRPA

Additional file 5. Titration of administered mAbs in the serum of the passively immunized mice. The concentration of administered PfCyRPA-specific mAbs SB1.6 and c12 in the circulation was estimated by indirect ELISA on day one and eight after injection. For the detection of PfCyRPA-specific mAbs and total circulating IgGs, plates were coated with N-CyRPA or goat anti-mouse IgG (γ-chain specific) mAb (M1397, Sigma), respectively. After blocking, plates were incubated with dilutions of individual mouse serum. An HRP-conjugated goat anti-mouse IgG (γ-chain specific) Ab (A3673, Sigma) was used as secondary antibody and TMB as substrate. Standard curves were generated from known dilutions of SB1.6 and c12 mAbs and fit using a 4-PL logistic equation. Concentration of circulating mAbs was calculated by interpolating the absorbance values for the test sera from the standard curves. Reported values are means of three mice per group ± SD.