Identification of the epitopes targeted both by FV2 and DBL4e specific binding inhibitory antibodies may serve as a tool to generate second-generation antigens by removing or masking redundant/ unwanted epitopes. We do not know how the immune response will be focused in humans upon immunization with DBL4e and the results from a phase 1 clinical trial in human volunteers with DBL4e will be decisive for further clinical development. Retrospectively, we would probably not have identified the binding inhibitory capacity of DBL4e specific antibodies had we only tested the antigen in rabbits and mice. This emphasises the value of testing different animal species as vaccination models, specifically for antigens like PfEMP-1 and EBA, which have evolved to generate variations in humoral immune responses. Plasmodium falciparum culture P. falciparum, FCR3 parasite (laboratory strain) were cultured in RPMI-1640 supplemented with 25 mmol/L sodium bicarbonate (Sigma-Aldrich), 0.125 mg/ml gentamycin, 0.125 mg/ml Albumax II (Invitrogen), 2% normal human serum and with 5% hematocrit of group 0+ human blood. To select for VAR2CSA expression, IE were repeatedly panned on BeWo-cells as described [38] . All isolates were mycoplasma negative and were regularly genotyped using nested GLURP and MSP-2 primers in a single PCR step.