Array-based ELISAs for high-throughput analysis of human cytokines
Array-based ELISAs for high-throughput analysis of human cytokines
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DOI：
10.2144/01311dd03
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发表时间：
2001-07-01
期刊：
影响因子：
2.7
通讯作者：
Burns, C
中科院分区：
文献类型：
--
作者：
Moody, MD;Van Arsdell, SW;Burns, C
在这份报告中，我们描述了一个小型阵列系统的发展，适用于高通量定量的蛋白质。该微型阵列是一种多重夹心ELISA，可从96孔板的每个孔中的单个样品中测量七种不同的人细胞因子TNF-α、IFN α、IFN γ、IL-1 α、IL-1 β、IL-6和IL-10的浓度。通过在96孔聚苯乙烯板的威尔斯孔底部以3 × 3模式点样单克隆抗体（mAb）来产生微型阵列。通过使用生物素化的mAb，然后加入链霉亲和素-辣根过氧化物酶（HRP）偶联物和荧光底物，检测阵列mAb捕获的细胞因子。通过用市售的CCD相机对整个板成像，在阵列中的每个点处测量底物的HRP催化氧化产生的光。在这里，我们证明了这些96孔板格式的微型阵列具有的性能特点，使它们适用于抗炎化合物的高通量筛选。
In this report, we describe the development of a mini-array system suitable for high-throughput quantification of proteins. This mini-array is a multiplexed, sandwich;vpe ELISA that measures the concentration of seven different human cytokines-TNF-alpha, IFN alpha, IFN gamma, IL-1 alpha, IL-1 beta, IL-6, and IL-10-from a single sample in each,well of a 96-well plate. The mini-array is produced by spotting monoclonal antibodies (mAbs) in a 3 x 3 pattern in the bottom of the wells of 96-well polystyrene plates. Cytokines that are captured by the arrayed mAbs are detected by using biotinylated mAbs, followed by the addition of a streptavidin-horseradish peroxidase (HRP) conjugate and a chemiluminescent substrate. The light producted from HRP-catalyzed oxidation of the substrate is measured at each spot in the array by imaging the entire plate with a commercially available CCD camera. Here, we demonstrate that these 96-well-plate format mini-arrays have performance characterstics that make them suitable for the high-throughput screening of anti-inflammatory compounds.