A rapid and scalable density gradient purification method for Plasmodium sporozoites

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

Research output: Contribution to journalArticlepeer-review

53 Scopus citations

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.

Original languageEnglish (US)
Article number421
JournalMalaria journal
Volume11
DOIs
StatePublished - 2012

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Infectious Diseases

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