Rabbits were immunized with exoantigens from the spent medium of Plasmodium falciparum cultures, and the resultant immunologic responses were studied by indirect fluorescent antibody (IFA), hemagglutination (HA), and two-dimensional crossed immunoelectrophoretic (IEP) techniques. By crossed IEP, three parasite antigens, identified and characterized as proteins lacking lipid and carbohydrate moieties, reacted with rabbit antiserum and human immune serum. Pre-immunization sera of the rabbits used in these experiments had 1:80 IFA titers against P. falciparum schizonts which were then boosted eightfold by immunization with parasite exoantigens, in contrast to IFA titers of <1:2 for coccidia- and Pasteurella-free rabbits. Experimental infections of coccidia- and Pasteurella-free rabbits with rabbit Eimeria spp. resulted in 1:80 anti-P. falciparum IFA titers, suggesting cross-reactivity of coccidial and plasmodial antigens. Post-immunization sera demonstrated extremely high HA titers against human erythrocytes, underscoring the potency of human blood components containing parasite antigens prepared from cultures. These results suggest that coccidia- and Pasteurella-free rabbits may be useful in the analysis of antigen of cultured P. falciparum.