Integrated Bioluminescent Immunoassays for High-Throughput Sampling and Continuous Monitoring of Cytokines,Analytical Chemistry

当前位置： X-MOL 学术 › Anal. Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Integrated Bioluminescent Immunoassays for High-Throughput Sampling and Continuous Monitoring of Cytokines
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-05-30 , DOI: 10.1021/acs.analchem.3c00745
Eva A van Aalen _{1,

2} , Bas J H M Rosier _{1,

2} , Tom Jansen _{1,

2} , Simone F A Wouters _{1,

2} , Robin T Vermathen _{1,

2} , Harmen J van der Veer _{1,

2} , José Yeste Lozano ₃ , Sheeza Mughal ₃ , Juan M Fernández-Costa ₃ , Javier Ramón-Azcón _{3,

4} , Jaap M J den Toonder _{2,

5} , Maarten Merkx _{1,

2}

Affiliation

Immunoassays show great potential for the detection of low levels of cytokines, due to their high sensitivity and excellent specificity. There is a particular demand for biosensors that enable both high-throughput screening and continuous monitoring of clinically relevant cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNFα). To this end, we here introduce a novel bioluminescent immunoassay based on the ratiometric plug-and-play immunodiagnostics (RAPPID) platform, with an improved intrinsic signal-to-background and an >80-fold increase in the luminescent signal. The new dRAPPID assay, comprising a dimeric protein G adapter connected via a semiflexible linker, was applied to detect the secretion of IL-6 by breast carcinoma cells upon TNFα stimulation and the production of low concentrations of IL-6 (∼18 pM) in an endotoxin-stimulated human 3D muscle tissue model. Moreover, we integrated the dRAPPID assay in a newly developed microfluidic device for the simultaneous and continuous monitoring of changes in IL-6 and TNFα in the low-nanomolar range. The luminescence-based read-out and the homogeneous nature of the dRAPPID platform allowed for detection with a simple measurement setup, consisting of a digital camera and a light-sealed box. This permits the usage of the continuous dRAPPID monitoring chip at the point of need, without the requirement for complex or expensive detection techniques.

中文翻译：

用于高通量采样和连续监测细胞因子的集成生物发光免疫分析

由于其高灵敏度和出色的特异性，免疫测定显示出检测低水平细胞因子的巨大潜力。对生物传感器有特殊需求，能够实现高通量筛选和连续监测临床相关细胞因子，如白细胞介素 6 (IL-6) 和肿瘤坏死因子-α (TNFα)。为此，我们在这里介绍了一种基于比例即插即用免疫诊断 (RAPPID) 平台的新型生物发光免疫测定，具有改进的内在信号背景和 >80 倍的发光信号增加。新的 dRAPPID 分析，包括通过半柔性接头连接的二聚体蛋白 G 接头，用于检测 TNFα 刺激后乳腺癌细胞分泌的 IL-6 和在内毒素刺激的人 3D 肌肉组织模型中产生低浓度 IL-6 (~18 pM)。此外，我们将 dRAPPID 测定集成到新开发的微流体装置中，用于同时和连续监测低纳摩尔范围内 IL-6 和 TNFα 的变化。基于发光的读出和 dRAPPID 平台的均匀性质允许通过简单的测量设置进行检测，包括数码相机和光密封盒。这允许在需要时使用连续的 dRAPPID 监控芯片，而不需要复杂或昂贵的检测技术。我们将 dRAPPID 测定整合到新开发的微流体装置中，用于同时和连续监测低纳摩尔范围内 IL-6 和 TNFα 的变化。基于发光的读出和 dRAPPID 平台的均匀性质允许通过简单的测量设置进行检测，包括数码相机和光密封盒。这允许在需要时使用连续的 dRAPPID 监控芯片，而不需要复杂或昂贵的检测技术。我们将 dRAPPID 测定整合到新开发的微流体装置中，用于同时和连续监测低纳摩尔范围内 IL-6 和 TNFα 的变化。基于发光的读出和 dRAPPID 平台的均匀性质允许通过简单的测量设置进行检测，包括数码相机和光密封盒。这允许在需要时使用连续的 dRAPPID 监控芯片，而不需要复杂或昂贵的检测技术。

更新日期：2023-05-30

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11