Danielli Amos, Porat Noga, Ehrlich Marcelo, Arie Ady
Department of Biomedical Engineering, Washington University in St. Louis, USA.
J Vis Exp. 2010 Jun 13(40):1935. doi: 10.3791/1935.
A magnetic modulation biosensing system (MMB) [1,2] rapidly and homogeneously detected biological targets at low concentrations without any washing or separation step. When the IL-8 target was present, a 'sandwich'-based assay attached magnetic beads with IL-8 capture antibody to streptavidin coupled fluorescent protein via the IL-8 target and a biotinylated IL-8 antibody. The magnetic beads are maneuvered into oscillatory motion by applying an alternating magnetic field gradient through two electromagnetic poles. The fluorescent proteins, which are attached to the magnetic beads are condensed into the detection area and their movement in and out of an orthogonal laser beam produces a periodic fluorescent signal that is demodulated using synchronous detection. The magnetic modulation biosensing system was previously used to detect the coding sequences of the non-structural Ibaraki virus protein 3 (NS3) complementary DNA (cDNA) [2]. The techniques that are demonstrated in this work for external manipulation and condensation of particles may be used for other applications, e.g. delivery of magnetically-coupled drugs in-vivo or enhancing the contrast for in-vivo imaging applications.
一种磁调制生物传感系统(MMB)[1,2]能够在无需任何洗涤或分离步骤的情况下,快速且均匀地检测低浓度生物靶标。当存在白细胞介素-8(IL-8)靶标时,基于“夹心”的检测方法通过IL-8靶标以及生物素化的IL-8抗体,将带有IL-8捕获抗体的磁珠与链霉亲和素偶联荧光蛋白相连。通过两个电磁极施加交变磁场梯度,使磁珠产生振荡运动。附着在磁珠上的荧光蛋白被聚集到检测区域,它们进出正交激光束的运动会产生周期性荧光信号,该信号通过同步检测进行解调。磁调制生物传感系统先前已用于检测茨城县病毒非结构蛋白3(NS3)互补DNA(cDNA)的编码序列[2]。这项工作中展示的用于粒子外部操控和聚集的技术可用于其他应用,例如体内磁耦合药物递送或增强体内成像应用的对比度。