A fully-automated, six-plex single molecule immunoassay for measuring cytokines in blood.
A fully-automated, six-plex single molecule immunoassay for measuring cytokines in blood.
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DOI：
10.1016/j.jim.2015.04.017
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发表时间：
2015-09
影响因子：
2.2
通讯作者：
Duffy, David C.
中科院分区：
文献类型：
--
作者：
Rivnak, Andrew J.;Rissin, David M.;Kan, Cheuk W.;Song, Linan;Fishburn, Matthew W.;Piech, Tomasz;Campbell, Todd G.;DuPont, Derek R.;Gardel, Melissa;Sullivan, Sean;Pink, Brian A.;Cabrera, Carlos G.;Fournier, David R.;Duffy, David C.
我们报告了一个系统和测定进行全自动测量的6种蛋白质同时与单分子的灵敏度。该系统将样本、试剂和耗材的处理与单分子阵列成像模块（Simoa）相结合，以实现具有高灵敏度（~fg/mL）、多路复用和全自动化的免疫测定。在该系统上开发了用于IL-6、TNF-α、GM-CSF、IL-10、IL-1β和IL-1α的6重细胞因子Simoa测定。测定的检测限在0.01-0.03 pg/mL范围内，Simoa信号的平均不精密度（CV）为4.2%。本试验用于测量克罗恩病（CD）患者血浆中、抗TNF-α抗体药物治疗前后以及1型糖尿病患者血清中这些细胞因子的浓度。使用全自动、多重Simoa测定法测定CD样本中TNF-α和IL-6的浓度与先前报告的手动、单重测定法具有良好的相关性。药物治疗导致CD患者血浆中所有6种细胞因子的平均浓度降低。与健康对照组相比，糖尿病患者中4种细胞因子的浓度显著较高。该系统可以实现对低丰度蛋白质生物标志物的广泛、多重测量。
We report a system and assay for performing fully-automated measurement of 6 proteins simultaneously with single molecule sensitivity. The system combines handling of samples, reagents, and consumables, with a module for imaging single molecule arrays (Simoa) to enable immunoassays that have high sensitivity (~fg/mL), are multiplexed, and are fully-automated. A 6-plex cytokine Simoa assay for IL-6, TNF-α, GM-CSF, IL-10, IL-1β, and IL-1α was developed on the system. The assays had limits of detection in the range 0.01–0.03 pg/mL, and the average imprecision (CV) of the Simoa signal was 4.2%. This assay was used to measure the concentrations of these cytokines in the plasma of patients with Crohn‘s Disease (CD), before and after treatment with anti-TNF-α antibody drugs, and in the serum of Type 1 diabetics. Concentrations of TNF-α and IL-6 in the CD samples determined using the fully-automated, multiplex Simoa assay had good correlation with the manual, single-plex assays previously reported. Drug treatment caused reductions in the mean concentration of all 6 cytokines in the plasma of CD patients. The concentrations of 4 cytokines were significantly higher in diabetics compared to healthy controls. The system could enable the widespread, multiplexed measurement of protein biomarkers with low abundance.
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影响因子：
2.2
作者：
Song, Linan;Hanlon, David W.;Duffy, David C.
通讯作者：
Duffy, David C.
影响因子：
9.3
作者：
Todd, John;Freese, Bob;Goix, Philippe
通讯作者：
Goix, Philippe
影响因子：
7.4
作者：
Rissin, David M.;Fournier, David R.;Piech, Tomasz;Kan, Cheuk W.;Campbell, Todd G.;Song, Linan;Chang, Lei;Rivnak, Andrew J.;Patel, Purvish P.;Provuncher, Gail K.;Ferrell, Evan P.;Howes, Stuart C.;Pink, Brian A.;Minnehan, Kaitlin A.;Wilson, David H.;Duffy, David C.
通讯作者：
Duffy, David C.
影响因子：
6.1
作者：
Kan, Cheuk W.;Rivnak, Andrew J.;Duffy, David C.
通讯作者：
Duffy, David C.
影响因子：
1.3
作者：
Kim, Jae Hyeon;Jin, Sang-Man;Lee, Myung-Shik
通讯作者：
Lee, Myung-Shik