Another cellulose membrane containing the seventeen peptides were prepared, blocked and probed with LmmAbB2D4 (10 μg/ml). As shown in Fig. 2B, the peptides recognized by LmmAbB2D4 were peptide 4 (QCTMDQGRLRCR), CAL-101 purchase peptide 7 (TCATDQGRLRCT), peptide 8 (HCFHDQGRVRCA), peptide 14 (HCTMDQGRLRCR) and peptide 15 (SCMLDQGRSRCR). Analysis of these sequences revealed no obvious homology between the mimotopes and the mut-II sequence. Based on the results of immunoassay with cellulose-bound peptides, the peptides (QCTMDQGRLRCR, TCATDQGRLRCT, HCFHDQGRVRCA and HCTMDQGRLRCR) were synthesized in a soluble form, trapped

in liposomes and used as immunogens in rabbits. One week after the sixth injection, sera from rabbits were tested in an indirect ELISA for their reactivity toward the peptides, the L. muta whole venom and the cognate mut-II protein. The sera from rabbits immunized with peptides show marginal reactivity against the peptides coated to plates, likely due to low adsorption of peptides to the microtiter plates (data MI-773 mouse not shown). However, ELISA reactivity was observed when the

antigens were L. muta crude venom and mut-II ( Fig. 3A and B). The strongest reactivity toward Mut-II was obtained with the serum of rabbits immunized with peptides TCATDQGRLRCT and QCTMDQGRLRCR ( Fig. 3B). The serum of rabbits immunized with the peptides HCFHDQGRVRCA and HCTMDQGRLRCR reacted poorly with the Mut-II protein, even lower that the serum of mock-liposomes immunized rabbits. The neutralizing properties of the anti-peptide antibodies raised in rabbits were assessed in vivo by testing the hemorrhagic inducing activity

of L. muta venom in animals immunized with the four target peptides. The rabbits immunized with the peptide-mimotopes TCATDQGRLRCT and QCTMDQGRLRCR were completely protected ( Fig. 4A and B). The rabbits immunized with the HCFHDQGRVRCA and HCTMDQGRLRCR peptides were partially protected (about 62% and 37% protection, respectively). The animal from the group that received the empty liposome (without peptides) as negative control liposome was not protected. Snake venoms are a cocktail of biologically active molecules, including toxins with enzymatic and non-enzymatic activities that have evolved to assist in the capture and digestion Bacterial neuraminidase of prey, as well as defense against predators. Human systemic envenomation is associated with a number of adverse effects, the nature and severity of which depends on the species of snake, the quantity of venom injected and the time period between envenomation and the administration of appropriate medical treatment. These effects may include paralysis, myolysis, blood coagulation disturbances and renal damage [7] and [41]. Bushmaster snake envenomation is characterized by serious hemorrhage, blood coagulation disorders, and renal failure; hemorrhage is the major complication resulting from envenomation by the pit vipers Bothrops and Lachesis snakebites [22].