The Plasmodium falciparum merozoite surface protein 1 (MSP1), MSP1-42 and MSP1-19 are protective malaria vaccines. MSP1-42 is cleaved to form MSP1-33 and MSP1-19. The role of MSP1-33 in immunity is unclear. We investigated the antibody responses to MSP1-33; and to MSP1-33Trunc, in which major conserved sequences were excised. While anti-MSP1-33 antibodies were subdominant in the anti-MSP1-42 responses, immunizations with MSP1-33 or MSP1-33Trunc induced high levels of antibodies reactive with MSP1-42 or whole merozoites. Anti-MSP1-33 and anti-MSP1-33Tunc antibodies crossreacted with both allelic forms of MSP1-42. Anti-MSP1-33 sera were ineffective in inhibiting parasite growth in vitro; but they significantly enhanced the activities of sub-optimal concentrations of the inhibitory anti-MSP1-42 sera. Thus, immunization strategies with MSP1-based vaccines may benefit from co-induction of anti-MSP1-33 responses to enhance efficacy and potency.