Plasmodium falciparum: isolation and purification of spontaneously released merozoites by nylon membrane sieves.

摘要: Abstract A new procedure for isolating spontaneously released merozoites from in vitro cultures of Plasmodium falciparum (FVO and FCB strains) is described. The mature forms relatively synchronous containing predominantly trophozoites few schizonts were concentrated with Plasmagel then incubated at 37 C, without adding fresh red blood cells, until matured into schizonts. Merozoites which subsequently harvested freed host cell material by low-speed centrifugations nylon membrane sieves (3- 1.2-μm pore size). From a culture about 5.2 × 109 mature-form parasites, total 10.7 during three consecutive harvests 69% these recovered after the isolation purification procedures. As demonstrated both light electron microscopy, most morphologically intact merozoite preparations free constituents. SDS-acrylamide gel electrophoresis confirmed absence also showed that had complex protein pattern apparent molecular weights between 225 15 kdaltons. Such purified will be invaluable malaria immunization studies, identification protective antigens P. falciparum, other immunological biochemical studies.