Ressine Anton, Ekström Simon, Marko-Varga György, Laurell Thomas
Department of Electrical Measurements, Lund Institute of Technology, Lund University, P.O. Box 118, S-221 00 Lund, Sweden.
Anal Chem. 2003 Dec 15;75(24):6968-74. doi: 10.1021/ac034425q.
The present work demonstrates the possibilities of using macroporous silicon as a substrate for highly sensitive protein chip applications. The formation of 3D porous silicon structures was performed by electrochemical dissolution of monocrystalline silicon. The fabricated macroporous silicon network has a rigid spongelike structure showing high uniformity and mechanical stability. The microfluidic properties of the substrates were found to be essential for a good bioassay performance. Small spot area, good spot reproducibility, and homogeneous spot profiles were demonstrated on the substrates for immobilized aRIgG. Water contact angles were measured on the porous surface and compared to that of planar silicon, silanized glass, and ordinary microscope glass slides. The effect of the porous surface on the performance of a model IgG-binding immunoassay is presented. aRIgG was microdispensed onto the chip surface forming a microarray of spots with high affinity for the target analyte. The dispensing was performed using an in-house-developed piezoelectric flow-through dispenser. Each spot was formed by a single droplet (100 pL) at each position. The macroporous silicon allowed a high-density microarraying with spot densities up to 4400 spots/cm2 in human plasma samples without cross-talk and consumption of only 0.6 pmol of antibodies/1-cm2 array. Antigen levels down to 70 pM were detected.
本研究展示了使用大孔硅作为高灵敏度蛋白质芯片应用底物的可能性。通过单晶硅的电化学溶解来形成三维多孔硅结构。所制备的大孔硅网络具有刚性海绵状结构,显示出高度的均匀性和机械稳定性。发现底物的微流体特性对于良好的生物测定性能至关重要。在固定抗人IgG的底物上展示了小斑点面积、良好的斑点重现性和均匀的斑点轮廓。在多孔表面测量了水接触角,并与平面硅、硅烷化玻璃和普通显微镜载玻片的水接触角进行了比较。展示了多孔表面对模型IgG结合免疫测定性能的影响。将抗人IgG微量分配到芯片表面,形成对目标分析物具有高亲和力的斑点微阵列。使用自行开发的压电流通式分配器进行分配。每个位置的每个斑点由单个液滴(100 pL)形成。大孔硅允许在人血浆样品中进行高密度微阵列,斑点密度高达4400个斑点/cm²,无串扰,每1 cm²阵列仅消耗0.6 pmol抗体。检测到低至70 pM的抗原水平。