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Title: A rapid and scalable density gradient purification method for Plasmodium sporozoites
Authors: Mark Kennedy
Matthew E. Fishbaugher
Ashley M. Vaughan
Rapatbhorn Patrapuvich
Rachasak Boonhok
Narathatai Yimamnuaychok
Nastaran Rezakhani
Peter Metzger
Marisa Ponpuak
Jetsumon Sattabongkot
Stefan H. Kappe
Jen Cc Hume
Scott E. Lindner
Seattle Biomedical Research Institute
Mahidol University
University of Washington, Seattle
Keywords: Immunology and Microbiology;Medicine
Issue Date: 19-Dec-2012
Citation: Malaria Journal. Vol.11, (2012)
Abstract: Background: Malaria remains a major human health problem, with no licensed vaccine currently available. Malaria infections initiate when infectious Plasmodium sporozoites are transmitted by Anopheline mosquitoes during their blood meal. Investigations of the malaria sporozoite are, therefore, of clear medical importance. However, sporozoites can only be produced in and isolated from mosquitoes, and their isolation results in large amounts of accompanying mosquito debris and contaminating microbes. Methods. Here is described a discontinuous density gradient purification method for Plasmodium sporozoites that maintains parasite infectivity in vitro and in vivo and greatly reduces mosquito and microbial contaminants. Results: This method provides clear advantages over previous approaches: it is rapid, requires no serum components, and can be scaled to purify > 10§ssup§7§esup§ sporozoites with minimal operator involvement. Moreover, it can be effectively applied to both human (Plasmodium falciparum, Plasmodium vivax) and rodent (Plasmodium yoelii) infective species with excellent recovery rates. Conclusions: This novel method effectively purifies viable malaria sporozoites by greatly reducing contaminating mosquito debris and microbial burdens associated with parasite isolation. Large-scale preparations of purified sporozoites will allow for enhanced in vitro infections, proteomics, and biochemical characterizations. In conjunction with aseptic mosquito rearing techniques, this purification technique will also support production of live attenuated sporozoites for vaccination. © 2012 Kennedy et al.; licensee BioMed Central Ltd.
ISSN: 14752875
Appears in Collections:Scopus 2011-2015

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