Analytical and preparative applications of magnetic split-flow thin fractionation on several ion-labeled red blood cells

Hweiyan Tsai,C Bor Fuh,Ying S Fang

doi:10.1186/1477-044x-4-6

Hweiyan Tsai, C Bor Fuh + Show 1 more

Open Access

https://doi.org/10.1186/1477-044x-4-6

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Abstract

BackgroundMagnetic Split-flow thin (SPLITT) fractionation is a newly developed technique for separating magnetically susceptible particles. Particles with different field-induced velocities can be separated into two fractions by adjusting applied magnetic forces and flow-rates at inlets and outlets.MethodsMagnetic particles, Dynabeads, were used to test this new approach of field-induced velocity for susceptibility determination using magnetic SF at different magnetic field intensities. Reference measurements of magnetic susceptibility were made using a superconducting quantum interference device (SQUID) magnetometer. Various ion-labeled red blood cells (RBC) were used to study susceptibility determination and throughput parameters for analytical and preparative applications of magnetic SPLITT fractionation (SF), respectively. Throughputs were studied at different sample concentrations, magnetic field intensities, and channel flow-rates.ResultsThe susceptibilities of Dynabeads determined by SPLITT fractionation (SF) were consistent with those of reference measurement using a superconducting quantum interference device (SQUID) magnetometer. Determined susceptibilities of ion-labeled RBC were consistent within 9.6% variations at two magnetic intensities and different flow-rates. The determined susceptibilities differed by 10% from referenced measurements. The minimum difference in magnetic susceptibility required for complete separation was about 5.0 × 10-6 [cgs]. Sample recoveries were higher than 92%. The throughput of magnetic SF was approximately 1.8 g/h using our experimental setup.ConclusionMagnetic SF can provide simple and economical determination of particle susceptibility. This technique also has great potential for cell separation and related analysis. Continuous separations of ion-labeled RBC using magnetic SF were successful over 4 hours. The throughput was increased by 18 folds versus early study. Sample recoveries were 93.1 ± 1.8% in triplicate experiments.

Highlights

Magnetic Split-flow thin (SPLITT) fractionation is a newly developed technique for separating magnetically susceptible particles
The determination of Dynabeads susceptibility Magnetic susceptibility determination of particles using analytical SPLITT fractionation (SF) was reported in the literature [5]
Determined susceptibilities of Dynabeads were consistent with relative standard deviation (RSD) within 5% variations and differed by a 5% range from reference measurements using superconducting quantum interference device (SQUID) for all measured conditions

Summary

Introduction

Magnetic Split-flow thin (SPLITT) fractionation is a newly developed technique for separating magnetically susceptible particles. Particles with different fieldinduced velocities can be separated into two fractions by adjusting applied magnetic forces and flow-rates at inlets and outlets. Split-flow thin (SPLITT) fractionation has become useful separation techniques for macromolecules, colloids, and particles [1-14]. In SPLITT fractionation (SF), thin (< 0.5 mm) channels without packing stationary phase are used and different forces are applied perpendicularly to the channel flow for separations. SF and field-flow fractionation (FFF) are close family of separation techniques for macromolecules, colloids, and particles [15-18]. Magnetic SF is a new member of SF family for separating magnetically susceptible colloids, and particles [2,3,5]

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