Tsai H Y, Hsu F H, Lin Y P, Bor Fuh C
School of Applied Chemistry, Chung Shan Medical University, Taichung, Taiwan.
J Chromatogr A. 2006 Oct 20;1130(2):227-31. doi: 10.1016/j.chroma.2006.05.045.
A separation method is reported for particle and biochemical analysis based on affinity interactions between particle surfaces under magnetic field. In this method, magnetic particles with immunoglobulin G (IgG) or streptavidin on the surface are flowed through a separation channel to form a deposition matrix for selectively capturing nonmagnetic analytes with protein A or biotin on the surface due to specific antigen (Ag)--antibody (Ab) interactions. This separation method was demonstrated using model reactions of IgG--protein A and streptavidin-biotin on particle surface. The features of this new separation method are (1) the deposited Ag-Ab complex can be examined and further analyzed under the microscope, (2) a kinetic study of complex binding is possible, and (3) the predeposited matrix can be formed selectively and changed easily. The detection limits were about 10(-11) g. The running time was less than 10 min. The selectivities of studied particles were 94% higher than those of label-controlled particles. This method extends the applications of analytical magnetapheresis to nonmagnetic particles. Preliminary study shows that this separation method has a great potential to provide a simple, fast, and selective analysis for particles, blood cells, and immunoassay related applications.
报道了一种基于磁场作用下颗粒表面亲和相互作用的颗粒和生化分析分离方法。在该方法中，表面带有免疫球蛋白G（IgG）或链霉亲和素的磁性颗粒流经分离通道，形成沉积基质，用于通过表面特异性抗原（Ag）-抗体（Ab）相互作用选择性捕获表面带有蛋白A或生物素的非磁性分析物。利用颗粒表面IgG-蛋白A和链霉亲和素-生物素的模型反应对该分离方法进行了验证。这种新分离方法的特点是：（1）沉积的Ag-Ab复合物可在显微镜下检查并进一步分析；（2）可以对复合物结合进行动力学研究；（3）预沉积基质可选择性形成且易于改变。检测限约为10^(-11) g。运行时间少于10分钟。所研究颗粒的选择性比标记对照颗粒高94%。该方法将分析磁泳的应用扩展到非磁性颗粒。初步研究表明，这种分离方法具有为颗粒、血细胞及免疫分析相关应用提供简单、快速和选择性分析的巨大潜力。
