Quantification of low-picomolar concentrations of TNF-α in serum using the dual-network microfluidic ELISA platform
Quantification of low-picomolar concentrations of TNF-α in serum using the dual-network microfluidic ELISA platform
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DOI：
10.1021/ac800427z
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发表时间：
2008-07-01
影响因子：
7.4
通讯作者：
Tabrizian, Maryam
中科院分区：
文献类型：
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作者：
Hermann, Marc;Veres, Teodor;Tabrizian, Maryam
对于研究和诊断目的，以成本和时间有效的方式检测低水平蛋白质的能力至关重要。在这项研究中，选择了细胞因子TNF-α（肿瘤坏死因子-α），一种广泛使用的炎症反应蛋白质指标，以证明作者开发的双网络微流控ELISA（酶联免疫吸附测定）平台能够快速定量血清中低浓度的这种生物标志物。通过对多个实验参数的优化，展示了该系统的可行性。满足基础和应用研究的要求，同时也与具有挑战性的临床应用相关，如败血症患者的诊断。对于浓度高达500 pg/mL的人TNF-α的定量，在培养基和血清中的灵敏度均为45 pg/mL（2.6 pM），内部和内部变异小于15%。在8个平行反应中完成测定的总时间小于1小时。
For both research and diagnostic purposes, the ability to detect low levels of proteins in a cost- and time-effective manner is essential. In this study, the cytokine TNF-alpha (tumor necrosis factor-alpha), a widely used protein indicator of inflammatory response, was chosen to demonstrate the ability of the dual-network microfluidic ELISA (enzyme-linked immunosorbent assay) platform developed by the authors to rapidly quantify low concentrations of this biomarker in serum. Through the optimization of several experimental parameters, the system was shown. to meet the requirements for fundamental and applied studies, while also being relevant for challenging clinical applications such as the diagnosis of septic patients. A sensitivity of 45 pg/mL (2.6 pM) in both culture medium and serum, with inter- and intravariations of less than 15%, was attained for the quantification of human TNF-alpha to a concentration of up to 500 pg/mL. The overall time for completion of the assay in eight parallel reactions was less than 1 h.