A Microfluidic Platform for High-Throughput Multiplexed Protein Quantitation

We present a high-throughput microfluidic platform capable of quantitating up to 384 biomarkers in 4 distinct samples by immunoassay. The microfluidic device contains 384 unit cells, which can be individually programmed with pairs of capture and detection antibody. Samples are quantitated in each unit cell by four independent MITOMI detection areas, allowing four samples to be analyzed in parallel for a total of 1,536 assays per device. We show that the device can be pre-assembled and stored for weeks at elevated temperature and we performed proof-of-concept experiments simultaneously quantitating IL-6, IL-1β, TNF-α, PSA, and GFP. Finally, we show that the platform can be used to identify functional antibody combinations by screening 64 antibody combinations requiring up to 384 unique assays per device.

本研究提出一种高通量微流控平台，可通过免疫测定（immunoassay）对4份不同样本中的至多384种生物标志物进行定量检测。该微流控器件包含384个微流控单元，每个单元可独立配置成对的捕获抗体与检测抗体。每个微流控单元通过四个独立的MITOMI检测区域完成样本定量，可并行分析4份样本，单台器件总计可完成1536次检测反应。本研究证实该器件可预组装并在高温环境下储存数周；同时开展了概念验证实验，同时定量检测白细胞介素6（IL-6）、白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）、前列腺特异性抗原（PSA）以及绿色荧光蛋白（GFP）。最后，本研究证明该平台可通过筛选64种抗体组合来鉴定功能性抗体配对，单台器件最多可完成384次独立检测。