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Plasmodium falciparum: isolation of intact and erythrocyte-free trophozoites from sorbitol lysates

Published online by Cambridge University Press:  06 April 2009

H. C. Hoppe
Affiliation:
Department of Biochemistry
J. Coetzee
Affiliation:
Electron Microscopy Unit, University of Pretoria, Pretoria 0002, Republic of South Africa
A. I. Louw
Affiliation:
Department of Biochemistry

Extract

An in vitro culture of Plasmodium falciparum (isolate FCUP-1/RSA) was arrested in the trophozoite stage with α-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.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1992

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