Hoyos M, Moore L R, McCloskey K E, Margel S, Zuberi M, Chalmers J J, Zborowski M
Ecole Supérieure de Physique et Chimie Industrielles, Laboratoire de Physique et Mécanique des Milieux Hétérogènes, UMR 7636 CNRS, Paris, France.
J Chromatogr A. 2000 Dec;903(1-2):99-116. doi: 10.1016/s0021-9673(00)00879-7.
Advantages of the continuous magnetic flow sorting for biomedical applications over current, batch-wise magnetic separations include high throughput and a potential for scale-up operations. A continuous magnetic sorting process has been developed based on the quadrupole magnetic field centered on an annular flow channel. The performance of the sorter has been described using the conceptual framework of split-flow thin (SPLITT) fractionation, a derivative of field-flow fractionation (FFF). To eliminate the variability inherent in working with a heterogenous cell population, we developed a set of monodisperse magnetic microspheres of a characteristic magnetization, and a magnetophoretic mobility, similar to those of the cells labeled with a magnetic colloid. The theory of the magnetic sorting process has been tested by injecting a suspension of the magnetic beads into the carrier fluid flowing through the sorter and by comparing the theoretical and experimental recovery versus total flow-rate profiles. The position of the recovery maxima along the total flow-rate axis was a function of the average bead magnetophoretic mobility and the magnetic field intensity. The theory has correctly predicted the position of the peak maxima on the total flow-rate axis and the dependence on the bead mobility and the field intensity, but has not correctly predicted the peak heights. The differences between the calculated and the measured peak heights were a function of the total flow-rate through the system, indicating a fluid-mechanical origin of the deviations from the theory (such as expected of the lift force effects in the system). The well-controlled elution studies using the monodisperse magnetic beads, and the SPLITT theory, provided us with a firm basis for the future sorter evaluation using cell mixtures.
与当前的分批式磁分离相比,连续磁流分选在生物医学应用中的优势包括高通量和扩大操作规模的潜力。基于以环形流道为中心的四极磁场,开发了一种连续磁分选工艺。已使用分流薄层(SPLITT)分级的概念框架来描述该分选器的性能,SPLITT分级是场流分级(FFF)的一个衍生方法。为了消除处理异质细胞群体时固有的变异性,我们开发了一组具有特征性磁化强度和磁泳迁移率的单分散磁性微球,其与用磁性胶体标记的细胞的磁化强度和磁泳迁移率相似。通过将磁珠悬浮液注入流经分选器的载液中,并比较理论和实验回收率与总流速曲线,对磁分选过程的理论进行了测试。回收率最大值沿总流速轴的位置是平均磁珠磁泳迁移率和磁场强度的函数。该理论正确地预测了总流速轴上峰值最大值的位置以及对磁珠迁移率和场强的依赖性,但没有正确预测峰值高度。计算出的和测量到的峰值高度之间的差异是通过系统的总流速的函数,这表明偏差源于流体力学(如系统中升力效应所预期的那样)。使用单分散磁珠进行的良好控制的洗脱研究以及SPLITT理论,为我们未来使用细胞混合物评估分选器提供了坚实的基础。