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Thread-Based Bioluminescent Sensor for Detecting Multiple Antibodies in a Single Drop of Whole Blood.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-05-22 , DOI: 10.1021/acssensors.0c00564
Kosuke Tomimuro 1 , Keisuke Tenda 1 , Yan Ni 2 , Yuki Hiruta 1 , Maarten Merkx 1, 2 , Daniel Citterio 1
Affiliation  

Antibodies are important biomarkers in clinical diagnostics in addition to being increasingly used for therapeutic purposes. Although numerous methods for their detection and quantification exist, they predominantly require benchtop instruments operated by specialists. To enable the detection of antibodies at point-of-care (POC), the development of simple and rapid assay methods independent of laboratory equipment is of high relevance. In this study, we demonstrate microfluidic thread-based analytical devices (μTADs) as a new platform for antibody detection by means of bioluminescence resonance energy-transfer (BRET) switching sensor proteins. The devices consist of vertically assembled layers including a blood separation membrane and a plastic film with a sewn-in cotton thread, onto which the BRET sensor proteins together with the substrate furimazine have been predeposited. In contrast to intensity-based signaling, the BRET mechanism enables time-independent, ratiometric readout of bioluminescence signals with a digital camera in a darkroom or a smartphone camera with a 3D-printed lens adapter. The device design allows spatially separated deposition of multiple bioluminescent proteins on a single sewn thread, enabling quantification of multiple antibodies in 5 μL of whole blood within 5 min. The bioluminescence response is independent of the applied sample volume within the range of 5–15 μL. Therefore, μTADs in combination with BRET-based sensor proteins represent user-friendly analytical tools for POC quantification of antibodies without any laboratory equipment in a finger prick (5 μL) of whole blood.

中文翻译:

基于线程的生物发光传感器,用于检测单滴全血中的多种抗体。

除了越来越多地用于治疗目的之外,抗体在临床诊断中也是重要的生物标志物。尽管存在多种检测和定量方法,但它们主要需要由专家操作的台式仪器。为了能够在即时医疗点(POC)上检测抗体,开发与实验室设备无关的简单快速的检测方法具有高度相关性。在这项研究中,我们演示了基于微流控线程的分析设备(μTADs)作为通过生物发光共振能量转移(BRET)转换传感器蛋白进行抗体检测的新平台。这些装置由垂直组装的层组成,包括血液分离膜和带有缝制棉线的塑料膜,BRET传感器蛋白与呋喃嗪底物一起预先沉积在其上。与基于强度的信号发送相反,BRET机制可通过暗室中的数码相机或带有3D打印镜头适配器的智能手机相机,以时间独立的方式按比例读出生物发光信号。该设备设计允许在单个缝合线上将多个生物发光蛋白在空间上分开沉积,从而能够在5分钟内定量5μL全血中的多种抗体。在5-15μL范围内,生物发光响应与所施加的样品量无关。因此,μTAD与基于BRET的传感器蛋白的结合代表了用户友好的分析工具,可用于对POC定量抗体的全血,而无需任何实验室设备即可刺穿手指(5μL)。
更新日期:2020-06-26
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