Interference in microsphere flow cytometric multiplexed immunoassays for human cytokine estimation
Interference in microsphere flow cytometric multiplexed immunoassays for human cytokine estimation
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DOI：
10.1016/j.cyto.2006.12.002
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发表时间：
2006-11-01
期刊：
影响因子：
3.8
通讯作者：
Hooper, W. Craig
中科院分区：
文献类型：
--
作者：
Phillips, Donald J.;League, Stacy C.;Hooper, W. Craig
本研究描述了多重微珠免疫测定中人细胞因子测量的阳性和阴性干扰。在30个正常人血浆样本中测量的IL-6和TNF-α值的显著差异是明显的，这取决于测量是使用2重测定还是包埋在8个或更多细胞因子抗体对的多重测定中，以及在3个不同供应商的试剂盒中。含有专有封闭成分的样品稀释剂显示出极大地影响测量的细胞因子值的结果。此外，从加标样品中回收的IL-6和TNF-α表明分别有10%和26%的样品受到内源性抗体、可溶性受体或抗细胞因子抗体的显著负干扰。虽然很明显，多重免疫测定对细胞因子谱分析有很大的希望，但仍有重要问题需要进一步研究。特别需要的是普遍优化的样品稀释剂，统一校准的标准与质量值，和内部测定控制，这将大大促进实验室内的准确性和精密度和实验室间比较的细胞因子测量。干扰的可能原因和补救措施进行了讨论。(c)2007爱思唯尔有限公司出版
The present study describes positive and negative interference of human cytokine measurement in multiplexed bead-based immuno-assays. Significant differences in measured IL-6 and TNF-alpha values in 30 normal human plasma samples were apparent depending on whether measurements were with a 2-plex assay or embedded in a multiplex of 8-or more cytokine antibody pairs, as well as among the kits of 3-different vendors. Sample diluents containing proprietary blocking ingredients were shown to greatly affect the outcome of measured cytokine values. Additionally, recovery of IL-6 and TNF-alpha from spiked samples suggests significant negative interference from either endogenous antibodies, soluble receptors or anti-cytokine antibodies in 10% and 26% of samples, respectively. While it is evident that multiplexed immunoassays hold great promise for cytokine profiling, there are still important issues needing further study. Especially needed are universally optimized sample diluents, uniformly calibrated standards with mass values, and internal assay controls, which should greatly facilitate intralaboratory accuracy and precision and interlaboratory comparisons of cytokine measurements. Possible causes of interference and remedies are discussed. (c) 2007 Published by Elsevier Ltd.