Detection of multiple proteins in an antibody-based protein microarray system
Detection of multiple proteins in an antibody-based protein microarray system
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DOI：
10.1016/s0022-1759(01)00394-5
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发表时间：
2001-09-01
影响因子：
2.2
通讯作者：
Huang, RP
中科院分区：
文献类型：
--
作者：
Huang, RP
描述了一种新的基于抗体的蛋白质阵列测定。该方法结合了酶联免疫吸附试验（ELISA）的特异性、增强化学发光（ECL）的灵敏性和微斑点法的高通量等优点。在该系统中，捕获蛋白，抗体或抗原以阵列形式点样到膜上。然后将生物样品与膜一起孵育。在样品中的抗原或抗体与其相应的靶标结合并洗去未结合的蛋白质后，将膜暴露于辣根过氧化物酶（HRP）缀合的抗体。最后用ECL系统对信号进行可视化。实验表明，使用这种新方法可以同时检测多种细胞因子和抗体。这个过程非常简单，不需要复杂的设备。这个概念应该能够扩展到开发一个高通量的蛋白质阵列系统。这种新方法的未来应用包括直接蛋白质表达谱分析、免疫疾病诊断和发现新的生物标志物。(C)2001 Elsevier Science B.V.保留所有权利。
A new antibody-based protein array assay is described. This assay combines the advantages of the specificity of enzyme-linked immunosorbent assays (ELISA), sensitivity of enhanced chemiluminescence (ECL) and high-throughput of microspot. In this system, the capture proteins, either antibodies or antigens are spotted onto membranes in an array format. Biological samples are then incubated with membranes. After antigens or antibodies in the samples bind to their corresponding targets and unbound proteins are washed away, the membranes are exposed to Horseradish Peroxidase (HRP)-conjugated antibody(ies). The signals are finally visualized with ECL system. Experiments demonstrate that multiple cytokines and antibodies can be simultaneously detected using this new approach. The procedure is so simple that no sophisticated equipment is required. The concept should be able to be extended to develop a high-throughput protein array system. Future applications of this new approach include direct protein expression profiling, immunological disease diagnostics and discovery of new biomarkers. (C) 2001 Elsevier Science B.V. All rights reserved.