Isolation of Intracellular Parasites (Plasmodium falciparum) from Culture Using Free-Flow Electrophoresis: Separation of the Free Parasites According to Stages

Abstract
Parasitized human erythrocytes were concentrated from continuous cultures of P. falciparum from 5-7% up to 80-95% using Plasmagel. After aggregation of the cells with phytohemagglutinin, the aggregated erythrocytes were fragmented by passing them, with minimal force, through successive nylon filters of decreasing pore size (100-3 .mu.m). The mixture of liberated, free parasites, intact erythrocytes and erythrocyte membrane vesicles was separated using free-flow electrophoresis. Most of the fractions containing free parasites did not show contamination with erythrocyte constituents as determined by light microscopy and EM, polyacrylamide gel electrophoresis and enzymatic analysis. The various stages of free parasites of P. falciparum exhibited different electrical surface charges. Rings and trophozoites were highly negatively charged whereas schizonts and, in particular, merozoites showed low negative charges. The various stages could be isolated separate from each other.

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