One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells

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One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells

Publisher
BioMed Central
Publication Date
Mar 17, 2004
Source
PMC
Keywords
Disciplines
  • Chemistry
  • Medicine
License
Unknown

Abstract

1475-2875-3-7.fm ral ss BioMed CentMalaria Journal Open AcceMethodology One-step concentration of malarial parasite-infected red blood cells and removal of contaminating white blood cells Dai Thi Xuan Trang1, Nguyen Tien Huy1,2, Tohru Kariu1,2, Kunihiko Tajima1 and Kaeko Kamei*1 Address: 1Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan and 2Venture Laboratory, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan Email: Dai Thi Xuan Trang - [email protected]; Nguyen Tien Huy - [email protected]; Tohru Kariu - [email protected]; Kunihiko Tajima - [email protected]; Kaeko Kamei* - [email protected] * Corresponding author Abstract Background: Isolation of a concentrated, living preparation of malarial parasite-infected red blood cells (PRBCs) that have low contamination of white blood cells (WBCs) facilitates research on the molecular, biochemical and immunological aspects of malarial parasites. This is currently carried out by a two-step method, including the concentration of PRBCs using density gradient centrifugation through Percoll or Nycodenz, followed by the removal of host WBCs using a cellulose powder column or a commercially available filtration unit. These two-step methods can help isolate sufficient PRBCs, but they are laborious. In this study, a simplified one-step procedure that takes advantage of the difference between diamagnetic low-spin oxyhaemoglobin and paramagnetic haemozoin (haem polymer) was described. The paramagnetic polymer is deposited in the food vacuoles of the parasite, allowing the use of magnetic separation to efficiently and rapidly concentrate PRBCs while removing contaminating host WBCs. Methods: The magnetic removal of WBCs using a commercial LD column (MACS) was evaluated as a new method for concentrating and purifying PRBCs. To compare this method with the two density gradient centrifugation methods using Percoll or Nycodenz, we analysed the

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