Plasmodium falciparum: isolation of intact and erythrocyte-free trophozoites from sorbitol lysates.
An in vitro culture of Plasmodium falciparum (isolate FCUP-1/RSA) was arrested in the trophozoite stage with alpha-difluoromethylornithine (DFMO), followed by sorbitol treatment to liberate intracellular parasites from infected erythrocytes. Most of the unlysed erythrocytes and ghost membranes were removed by filtration, after agglutination with anti-erythrocyte monoclonal antibodies or wheat-germ agglutinin. A highly purified parasite preparation was obtained after chromatography on an immuno-affinity column consisting of polystyrene particles to which anti-erythrocyte monoclonal antibodies had been adsorbed. Isolated parasites were free of surrounding erythrocyte membranes and structurally intact as assessed by transmission electron microscopy, SDS-PAGE and ELISA.
Hoppe
H C
HC
Department of Biochemistry, University of Pretoria, Republic of South Africa.
Coetzee
J
J
Louw
A I
AI
D000371:Agglutination; D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000187:drug effects / Q000469:parasitology*; D005374:Filtration; D006801:Humans; D051379:Mice; D008854:Microscopy, Electron; D010963:Plasmodium falciparum / Q000737:chemistry / Q000302:isolation & purification* / Q000648:ultrastructure; D013012:Sorbitol / Q000494:pharmacology*
D013012:Sorbitol