Parameterization of high magnetic field gradient fractionation columns for applications with Plasmodium falciparum infected human erythrocytes.
BACKGROUND
Magnetic fractionation of erythrocytes infected with Plasmodium falciparum has several research uses including enrichment of infected cells from parasite cultures or enhanced detection of P. falciparum gametocytes. The aim of the present study was to quantitatively characterize the magnetic fractionation process and thus enable optimization of protocols developed for specific uses.


METHODS
Synchronized cultures of P. falciparum parasites incubated with human erythrocytes were magnetically fractionated with commercially available columns. The timing of the fractionation experiments was such that the parasites were in second half of their erythrocytic life cycle with parasite densities ranging from 1 to 9%. Fractionations were carried out in a single pass through the columns. Cells were enumerated and differentiated in the initial samples as well as in the positive and negative fractions. The capture of cells by the fractionation column was described by a saturation binding model.


RESULTS
The magnetic binding affinity to the column matrix was approximately 350 times greater for infected cells compared with uninfected cells. The purity of infected cells in the captured fraction was generally >80% but decreased rapidly (to less than 50%) when the number of infected cells that passed through the column was substantially decreased (to less than 9 +/- 5 x 105 cells). The distribution of captured parasite developmental stages shifted to mature stages as the number of infected cells in the initial samples and flow rate increased. The relationship between the yield of infected cells in the captured fraction and flow rate of cells conformed to a complementary cumulative log-normal equation with flow rates >1.6 x 105 cells per second resulting in yields <50%.


CONCLUSIONS
A detailed quantitative analysis of a batchwise magnetic fractionation process for malaria infected erythrocytes using high gradient magnetic fractionation columns was performed. The models applied in this study allow the prediction of capture efficiency if the initial infected cell concentration and the flow rate are known.
Karl
Stephan
S
School of Physics, M013, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia.
Davis
Timothy M E
TM
St Pierre
Tim G
TG
D002458:Cell Fractionation / Q000379:methods*; D004903:Erythrocyte Aggregation; D004912:Erythrocytes / Q000166:cytology / Q000469:parasitology*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D008280:Magnetics / Q000295:instrumentation*; D016778:Malaria, Falciparum / Q000097:blood* / Q000469:parasitology; D009996:Osmotic Fragility; D018512:Parasitemia; D010963:Plasmodium falciparum / Q000254:growth & development / Q000302:isolation & purification*; D053058:Trophozoites / Q000502:physiology
D005456:Fluorescent Dyes