High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays
Danika L Hill, Emily M Eriksson, Louis Schofield
Journal of Visualized Experiments | JOURNAL OF VISUALIZED EXPERIMENTS | Published : 2014
Plasmodium falciparum merozoite antigens are under development as potential malaria vaccines. One aspect of immunity against malaria is the removal of free merozoites from the blood by phagocytic cells. However assessing the functional efficacy of merozoite specific opsonizing antibodies is challenging due to the short half-life of merozoites and the variability of primary phagocytic cells. Described in detail herein is a method for generating viable merozoites using the E64 protease inhibitor, and an assay of merozoite opsonin-dependent phagocytosis using the pro-monocytic cell line THP-1. E64 prevents schizont rupture while allowing the development of merozoites which are released by filtr..View full abstract
Awarded by National Health and Medical Research Council
Awarded by National Institutes of Health
Awarded by NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
The authors wish to acknowledge the children and adult plasma donors, and the staff at the Papua New Guinea Institute of Medical Research. The authors would like to thank Amandine B Carmagnac, Catherine Q Nie, Danny W Wilson, Ivo Mueller and Diana S Hansen for their contribution to development of this technique, and thank the Australian Red cross for blood and serum packs. This work was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS. This work was supported by National Health and Medical Research Council grants # 1031212 and # 637406, and National Institutes of Health grant # AI089686.