Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines
Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines
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DOI：
10.1016/j.jim.2003.10.011
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发表时间：
2004-01-01
影响因子：
2.2
通讯作者：
Remick, D
中科院分区：
文献类型：
--
作者：
Copeland, S;Siddiqui, J;Remick, D
许多实验室希望以准确、可重复和快速的方式测量细胞因子。基于与传统ELISA相同的技术，已经开发了用于测量细胞因子的基于抗体的膜阵列。本研究的目的是比较与传统的ELISA方法，从膜阵列技术，一种低成本的蛋白质组学的形式所获得的结果。用活细菌（大肠杆菌或金黄色葡萄球菌）刺激稀释的人全血，或通过ELISA和膜蛋白阵列测量LPS和细胞因子。在通过ELISA测量的16种细胞因子中，仅IFN-γ低于检测水平。其他15种细胞因子的浓度高达几千皮克/毫升。在通过膜蛋白阵列测量的20种细胞因子中，仅可检测到3种（IL-6、IL-8和MIP-1 β）。此外，膜蛋白阵列没有检测到来自LPS刺激的血液的TNF-α。这些结果表明，低成本的膜蛋白阵列可能缺乏足够的灵敏度，以充分检测复杂的生物流体，如人血浆中的细胞因子水平。(C)2003 Elsevier B. V.保留所有权利。
Many labs wish to measure cytokines in an accurate, reproducible, and rapid manner. An antibody-based membrane array for measuring cytokines has been developed based on the same technology as the traditional ELISA. The aim of this study was to compare results obtained with the traditional ELISA method with those from the membrane array technology, a form of low-cost proteomics. Diluted human whole blood was stimulated with live bacteria (Escherichia coli, or Staphylococous aureus), or LPS and cytokines were measured both by ELISA and the membrane protein array. Of the 16 cytokines measured via ELISA, only IFN-gamma was below detection level. The other 15 cytokines were present in concentrations up to several thousand picograms/ml. Of the 20 cytokines measured via membrane protein array, only 3 could be detected (IL-6, IL-8 and MIP-1beta). Additionally, the membrane protein array did not detect TNF-alpha from the LPS-stimulated blood. These results indicate that the low-cost membrane protein array may lack sufficient sensitivity to adequately detect cytokines levels in complex biological fluids such as human plasma. (C) 2003 Elsevier B.V. All rights reserved.