OBJECTIVE To prepare and characterize the monoclonal antibodies (mAbs) against erythrocyte-binding antigen 175 of Plasmodium falciparum (EBA-175). METHODS BALB/ c mice were immunized with purified recombinant EBA-175 and mAbs against EBA-175 were prepared by means of hybridoma technique. Enzyme-linked immunosorbent assay (ELISA) and Western blotting were employed for characterization of the mAbs. RESULTS Six McAbs against EBA-175 antigen were obtained, 5 of which were identified as IgG1 and one as IgG2a. The titer of these aAbs was 1:12,800 to 1:25,600 in the ascites and 1:256 to 1:512 in supernatant, and ELISA demonstrated specific binding of the 4 mAbs (1F3, 2H5, 4A1 and 4H9)with Plasmodium falciparum. Three of these mAbs recognized the protein of EBA-175 as shown by Western blotting. CONCLUSIONS Six hybridoma cell lines secreting the mAbs against EBA175 with high specificity are successfully established.