Single molecule array (Simoa) assay with optimal antibody pairs for cytokine detection in human serum samples
Single molecule array (Simoa) assay with optimal antibody pairs for cytokine detection in human serum samples
复制标题
DOI：
10.1039/c5an01238d
复制
发表时间：
2015-01-01
期刊：
影响因子：
4.2
通讯作者：
Walt, David R.
中科院分区：
文献类型：
--
作者：
Wu, Danlu;Milutinovic, Milena Dumont;Walt, David R.
体液中细胞因子的浓度反映了患者的生理或病理状态，可用于预后、疾病诊断或监测治疗效果。然而，在健康或亚健康个体的体液中，许多细胞因子的浓度接近或低于当前方法的检测极限。在这里，我们选择抗体对用于10种细胞因子的单分子阵列（Simoa）检测，包括GM-CSF、tnf - α、ifn - γ、IL-1 β、IL-2、IL-4、IL-5、IL-6、IL-7和IL-10。所获得的检出限（lod）低至90 aM-6 fM。这些检测允许检测健康人血清样品中的细胞因子，其水平明显低于常规ELISA检测的检测限。我们提供了详细的抗体对信息以及健康人血清中10种细胞因子的浓度谱，为进一步的超灵敏免疫测定发展和未来的临床应用提供参考数据。
Concentrations of cytokines in bodily fluids reflect the physiological or pathological state of the patient and can be used for prognosis, disease diagnosis or for monitoring therapeutic efficacy. However, in the bodily fluids of healthy or sub-healthy individuals, many cytokines are present at concentrations that are near or below the detection limits of current methods. Here we selected antibody pairs to be employed in the single molecule array (Simoa) assay for ten cytokines including GM-CSF, TNF-alpha, IFN-gamma, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-7, and IL-10. The limits of detection (LODs) obtained were as low as 90 aM-6 fM. These assays allow detection of cytokines in healthy human serum samples at levels significantly below the detection limits of conventional ELISA assays. We provide detailed antibody pair information as well as the concentration profiles of ten cytokines in healthy human serum to serve as reference data for further ultrasensitive immunoassay development and future clinical applications.