School of Pharmaceutical Science & Technology, Tianjin University, Tianjin, China.
J Chromatogr A. 2011 Jun 24;1218(25):3908-14. doi: 10.1016/j.chroma.2011.04.065. Epub 2011 May 6.
Although magnetic field-flow fractionation (MgFFF) is emerging as a promising technique for characterizing magnetic particles, it still suffers from limitations such as low separation efficiency due to irreversible adsorption of magnetic particles on separation channel. Here we report a novel approach based on the use of a cyclic magnetic field to overcome the particle entrapment in MgFFF. This cyclic field is generated by rotating a magnet on the top of the spiral separation channel so that magnetic and opposing gravitational forces alternately act on the magnetic particles suspended in the fluid flow. As a result, the particles migrate transversely between the channel walls and their adsorption at internal channel surface is prevented due to short residence time which is controlled by the rotation frequency. With recycling of the catch-release process, the particles follow saw-tooth-like downstream migration trajectories and exit the separation channel at velocities corresponding to their sedimentation coefficients. A retention model has been developed on the basis of the combined effects of magnetic, gravitational fields and hydrodynamic flow on particle migration. Two types of core-shell structured magnetic microspheres with diameters of 6.04- and 9.40-μm were synthesized and used as standard particles to test the proposed retention theory under varying conditions. The retention ratios of these two types of particles were measured as a function of magnet rotation frequency, the gap between the magnet and separation channel, carrier flow rate, and sample loading. The data obtained confirm that optimum separation of magnetic particles with improved separation efficiency can be achieved by tuning rotation frequency, magnetic field gradient, and carrier flow rate. In view of the widespread applications of magnetic microspheres in separation of biological molecules, virus, and cells, this new method might be extended to separate magnetically labeled proteins or organisms for multiplex analyte identification and purification.
尽管磁场流动分馏(MgFFF)作为一种有前途的技术正在出现,用于表征磁性颗粒,但它仍然存在一些限制,例如由于磁性颗粒不可逆地吸附在分离通道上,导致分离效率低。在这里,我们报告了一种基于使用循环磁场来克服 MgFFF 中颗粒捕获的新方法。这种循环磁场是通过在螺旋分离通道顶部旋转磁铁产生的,使得磁场和反向重力交替作用于悬浮在流体中的磁性颗粒。结果,颗粒在通道壁之间横向迁移,由于停留时间短(由旋转频率控制),其在内部通道表面的吸附被阻止。通过回收捕获-释放过程,颗粒遵循锯齿状的下游迁移轨迹,并以与它们的沉降系数相对应的速度从分离通道中流出。基于磁场、重力场和流体力对颗粒迁移的综合影响,建立了保留模型。合成了两种具有 6.04-和 9.40-μm 直径的核壳结构磁性微球作为标准颗粒,在不同条件下测试所提出的保留理论。作为功能测试,测量了这两种类型的颗粒在不同磁铁旋转频率、磁铁与分离通道之间的间隙、载体流速和样品加载条件下的保留率。获得的数据证实,通过调整旋转频率、磁场梯度和载体流速,可以实现磁性颗粒的最佳分离,提高分离效率。鉴于磁性微球在分离生物分子、病毒和细胞方面的广泛应用,这种新方法可能会扩展到分离磁性标记的蛋白质或生物体,用于多分析物识别和纯化。