High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA)
High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA)
复制标题
DOI：
10.2144/99274rr01
复制
发表时间：
1999-10-01
期刊：
影响因子：
2.7
通讯作者：
Eggers, M
中科院分区：
文献类型：
--
作者：
Mendoza, LG;McQuary, P;Eggers, M
新一代生物芯片被描述为能够支持高通量（HT）、多重酶联免疫吸附测定（ELISA）。这些生物芯片由光学平坦的玻璃板组成，所述玻璃板含有由封闭的疏水Teflon（R）掩模形成的96个威尔斯孔。每个孔和板的足迹尺寸与标准微孔板的足迹尺寸精确匹配。每个孔含有四个相同的36元件阵列（每孔144个元件），其包含8种不同的抗原和标记蛋白。阵列由连接到精确、高速、X-Y-Z机器人的定制连续流动、基于毛细管的打印头形成。单个机器人的阵列打印能力超过每天20 000个阵列。使用定制的高分辨率扫描电荷耦合器件（CCD）检测器对阵列进行定量成像，成像通量为每30秒96个阵列。使用这种新方法，使用标准ELISA技术单独和集体检测阵列抗原。实验表明，在玻璃基片上阵列的蛋白质抗原的特异性多重检测是可行的。由于开放的微阵列架构，96孔微阵列格式与自动化机器人系统兼容，并支持低成本，高度平行的测定格式。这种新的高通量筛选（HTS）格式的未来应用包括直接细胞蛋白质表达谱多重测定，用于检测感染因子和癌症诊断。
A new generation biochip is described as capable of supporting high-throughput (HT), multiplexed enzyme-linked immunosorbent assays (ELISAs). These biochips consist of an optically flat, glass plate containing 96 wells formed by an enclosing hydrophobic Teflon(R) mask. The footprint dimensions of each well and the plate precisely match those of a standard microplate. Each well contains four identical 36-element arrays (144 elements per well) comprising 8 different antigens and a marker protein. Arrays are formed by a custom continuous flow, capillary-based print head attached to a precise, high-speed, X-Y-Z robot. The array printing capacity of a single robot exceeds 20 000 arrays per day. Arrays are quantitatively imaged using a custom, high-resolution, scanning charge-coupled device (CCD) detector with an imaging throughput of 96 arrays every 30 s. Using this new process, arrayed antigens were individually and collectively detected using standard ELISA techniques. Experiments demonstrate that specific multiplex detection of protein antigens arrayed on a glass substrate is feasible. Because of the open microarray architecture, the 96-well microarray format is compatible with automated robotic systems and supports a low-cost, highly parallel assay format Future applications of this new high-throughput screening (HTS)format include direct cellular protein expression profiling multiplexed assays for detection of infectious agents and cancer diagnostics.