Preparation of Nanoliposomes Incorporated Leishmania donovani Antigens

Abstract

This study was designed to incorporate leishmania donovani antigens in nanoliposomes prepared by size exclusion (using Sephadex G25) and organic solvent (using Chloroform). Lipids mixture of 4Mm Phosphatidylcholine, 2.2mM Cholesterol and 0.55mM Phosphatidylethanolamine in a ratio of 7:2:1 was depended in two nanoliposome preparation methods. Physio-chemical characterizations of prepared nanoliposomes was performed by using Scanning Electron Microscope (SEM ), Fourier transform infrared spectroscopy (FTIR) and  Zeta Potential assays to determine the size, morphology, chemical active group and charge . Parasite reactivation was carried out when inoculated into RPMI and incubated at 23 ̊ C for 4 days. Soluble Leishmania Antigenes (SLAs) were extracted from the promastigotes ghost membrane after fourth passages of subculturing in SNB9. The extracted SLAs were entrapped in prepared. The percentage of nanoliposomes entrapment efficiency (EE) was 62 and 50 of SLAs for chloroform and Sephadex G25 methods, respectively. Moreover, stability of SLAs entrapped nanoliposomes at 4 and 37 ̊C, as storage temperature, was examined. The stability at 4 °C showed decreasing in EE to 32 and 16 %, while stability at 37 °C revealed decreasing in EE to 16 and 8 % within 12 days of storage for nanoliposomes prepared in both methods, respectively.