Separation of Individual Stages of Trypanosoma cruzi Grown in Cell Culture by Continuous Free‐Flow Electrophoresis

Publisher Website
Google Scholar
Abstract
The separation of extracellular protozoan parasites from host cells based on a difference in surface charge is described. With [the human parasite] T. cruzi no method exists for the isolation of pure parasite stages from heterogeneous mixtures. Studies on the electrophoresis of mixed stage populations confirm significant surface charge density differences exist among epimastigotes, trypomastigotes and amastigotes. In ascending order of electronegativity, amastigotes have the lowest charge density, trypomastigotes next, followed by epimastigotes. A technique was developed for the separation of purified populations of parasites based on these charge differences using a continuous free-flow electrophoresis apparatus. The separated populations are morphologically intact and maintain their infectivity to mice. This separation method is applicable for preparative and analytical isolation of pure stages of T. cruzi for biochemical and immunological studies.