Effects of serum and plasma matrices on multiplex immunoassays.
Effects of serum and plasma matrices on multiplex immunoassays.
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DOI：
10.1007/s12026-014-8491-6
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发表时间：
2014-05
影响因子：
4.4
通讯作者：
Maecker, Holden T.
中科院分区：
文献类型：
--
作者：
Rosenberg-Hasson, Yael;Hansmann, Leo;Liedtke, Michaela;Herschmann, Iris;Maecker, Holden T.
可溶性细胞因子的多路荧光或电化学发光免疫分析通常在人血清或血浆中进行，以寻找疾病生物标志物并以简单和微创的方式监测免疫系统。由于基质（血清或血浆）的复杂性以及许多细胞因子接近检测极限的存在，这些检测提供了挑战。在这里，我们比较了匹配的血清和血浆样本的读数，它们通常是相关的。然而，一组细胞因子通常在血清中有较高的水平，非特异性背景在血清中比在血浆中显著增加。可能是由于这种非特异性背景，低丰度细胞因子的疾病相关减少有时可以在血浆中检测到，但在血清中检测不到。我们进一步表明，通过尖峰恢复实验，血清和血浆都抑制许多细胞因子的读数，在供体之间有一些差异，但在许多情况下，血清比血浆产生更大的抑制作用。来自不同供应商的标准稀释剂可以在不同程度上部分逆转这种抑制作用。样品的稀释也可以部分克服基质的抑制作用。我们还表明，稀释是非线性的，对各种细胞因子的影响是不同的。总之，这些数据表明：(1)血浆是检测某些低丰度细胞因子变化的更敏感的基质；(2)计算血清或血浆基质中的浓度本身是不准确的；(3)不能假设样品的稀释是线性的，即所有的比较都需要在相似稀释的样品之间进行。
Multiplexed fluorescence or electrochemiluminescence immunoassays of soluble cytokines are commonly performed in the context of human serum or plasma, to look for disease biomarkers and to monitor the immune system in a simple and minimally invasive way. These assays provide challenges due to the complexities of the matrix (serum or plasma) and the presence of many cytokines near the limit of detection of the assay. Here, we compare the readout of matched serum and plasma samples, which are generally correlated. However, a subset of cytokines usually have higher levels in serum, and the non-specific background is significantly increased in serum versus plasma. Presumably as a result of this non-specific background, disease-related decreases in low-abundance cytokines can sometimes be detected in plasma but not in serum. We further show, through spike recovery experiments, that both serum and plasma inhibit the readout of many cytokines, with some variability between donors, but with serum causing greater inhibition than plasma in many cases. Standard diluents from different vendors can partially reverse this inhibition to varying degrees. Dilution of samples can also partly overcome the inhibitory effect of the matrix. We also show that dilution is nonlinear and differentially affects various cytokines. Together, these data argue that (1) plasma is a more sensitive matrix for detecting changes in certain low-abundance cytokines; (2) calculation of concentrations in serum or plasma matrices is inherently inaccurate; and (3) dilution of samples should not be assumed to be linear, i.e., all comparisons need to be made among similarly diluted samples.
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影响因子：
2.2
作者：
Dossus, Laure;Becker, Susen;Rinaldi, Sabina
通讯作者：
Rinaldi, Sabina
DOI：
10.1158/1055-9965.epi-11-0221
发表时间：
2011-09
期刊：
Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology
影响因子：
--
作者：
Chaturvedi AK;Kemp TJ;Pfeiffer RM;Biancotto A;Williams M;Munuo S;Purdue MP;Hsing AW;Pinto L;McCoy JP;Hildesheim A
通讯作者：
Hildesheim A
影响因子：
3.8
作者：
Phillips, Donald J.;League, Stacy C.;Hooper, W. Craig
通讯作者：
Hooper, W. Craig
影响因子：
3.8
作者：
Wong, Hui-Lee;Pfeiffer, Ruth M.;Rabkin, Charles S.
通讯作者：
Rabkin, Charles S.
DOI：
10.1128/cdli.11.2.325-329.2004
发表时间：
2004-03-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Martins, TB;Pasi, BM;Hill, HR
通讯作者：
Hill, HR