Tang Chi K, Vaze Abhay, Shen Min, Rusling James F
Department of Chemistry, University of Connecticut, Storrs, Connecticut 06269, United States.
Department of Chemistry, University of Connecticut, Storrs, Connecticut 06269, United States; Institute of Materials Science, University of Connecticut, Storrs, Connecticut 06269, United States; Department of Surgery and Neag Cancer Center, University of Connecticut Health Center, Farmington, Connecticut 06032, United States; School of Chemistry, National University of Ireland at Galway, Galway, Ireland.
ACS Sens. 2016 Aug 26;1(8):1036-1043. doi: 10.1021/acssensors.6b00256. Epub 2016 Jul 21.
Microchip-based microfluidic electrochemical arrays hold great promise for fast, high-throughput multiplexed detection of cancer biomarker proteins at low cost per assay using relatively simple instrumentation. Here we describe an inexpensive high-throughput electrochemical array featuring 32 individually addressable microelectrodes that is further multiplexed with an 8-port manifold to provide 256 sensors. The gold electrode arrays were fabricated by wet-etching commercial gold compact discs (CD-R) followed by patterned insulation. A print-and-peel method was used to create sub-microliter hydrophobic wells surrounding each sensor to eliminate cross contamination during immobilization of capture antibodies. High-throughput analyses were realized using eight 32-sensor immunoarrays connected to the miniaturized 8-port manifold, allowing 256 measurements in <1 h. This system was used to determine prostate cancer biomarker proteins prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), interleukin-6 (IL-6), and platelet factor-4 (PF-4) in serum. Clinically relevant detection limits (0.05 to 2 pg mL) and 5-decade dynamic ranges (sub pg mL to well above ng mL) were achieved for these proteins utilizing precapture of analyte proteins on magnetic nanoparticles decorated with enzyme labels and antibodies.
基于微芯片的微流控电化学阵列有望以相对简单的仪器设备，以较低的单次检测成本快速、高通量地多重检测癌症生物标志物蛋白。在此，我们描述了一种低成本的高通量电化学阵列，它具有32个可单独寻址的微电极，并通过一个8端口流路进行进一步多重化，从而提供256个传感器。金电极阵列通过对商用金光盘（CD-R）进行湿法蚀刻，然后进行图案化绝缘来制造。采用打印-剥离法在每个传感器周围创建亚微升疏水孔，以消除捕获抗体固定过程中的交叉污染。使用连接到小型化8端口流路的八个32传感器免疫阵列实现了高通量分析，可在不到1小时内进行256次测量。该系统用于测定血清中的前列腺癌生物标志物蛋白前列腺特异性抗原（PSA）、前列腺特异性膜抗原（PSMA）、白细胞介素-6（IL-6）和血小板因子-4（PF-4）。利用在装饰有酶标记和抗体的磁性纳米颗粒上预捕获分析物蛋白，实现了这些蛋白的临床相关检测限（0.05至2 pg/mL）和5个数量级的动态范围（亚pg/mL至远高于ng/mL）。
