Liposomes have long been used as models for lipid membranes, and for the reconstitution of single or multiple proteins. In addition to that, liposomes have an adjuvant activity in vaccines against protozoan or bacterial organisms. Thus, the main objective of the present study was to obtain a crude extract of detergent-solubilized proteins of L. amazonensis amastigotes and reconstitute them into liposomes. Neutral and zwiterionic detergents were less efficient than an ionic detergent. In order to obtain efficient solubilization using only SDS, the effects of detergent and protein concentration and incubation time were studied. The maximum solubilized proteins were obtained instantaneously using a ratio of 0.5 mg/ml of protein to 0.1% (w/v) detergent at 4oC. With the objective of preparing proteoliposomes, different mixtures of lipids (DPPC or DPPS alone or both phospholipids with or without cholesterol, in different proportions) and an SDS-solubilized proteins extract were used and an optimization of the incorporation of protein was obtained using the mixtures of DPPC, DPPS and cholesterol, at weight ratios of 5:1:4 and 0.5 mg/ml of protein. The incorporation of proteins into liposomes by the co-solubilization method requires the complete removal of the detergent. In this work, SDS removal was achieved by batch hydrophobic adsorption with the Calbiosorb resin giving a SDS-free proteoliposome. In our system, the reconstitution of the solubilized proteins was around 60%.  The incorporation of multiple parasite proteins results in a vesicular diameter of proteoliposomes of about 140 nm presenting a final lipid weight ratio for DPPC, DPPS and cholesterol of 1:1:5, with high stability. The detergent-solubilized proteins of L. amazonensis amastigotes present in the proteoliposome, when analyzed by SDS-PAGE, shows a wide range of parasite-incorporated proteins. BALB/c mice inoculated with these proteoliposomes were able to produce antibodies against the proteins reconstituted in DPPC:DPPS:cholesterol-liposomes and were partially resistant to infection with L. amazonensis promastigotes. These results indicate that this system can be used as a possible vaccine against L. amazonensis.  Supported by: CNPq, CAPES and FAPESP.