Issue 12, 2023

Highly parallel, wash-free, and ultrasensitive centrifugal droplet digital protein detection in sub-microliter blood

Abstract

The ability to efficiently detect low-abundance protein biomarkers in tiny blood samples is a significant challenge in clinical and laboratory settings. Currently, high-sensitivity approaches require specialized instrumentation, involve multiple washing steps, and lack the ability to parallelize, preventing their widespread implementation. Herein, we developed a parallelized, wash-free, and ultrasensitive centrifugal droplet digital protein detection (CDPro) technology that achieves a femtomolar limit of detection (LoD) of target proteins with sub-microliters of plasma. The CDPro combines two techniques, namely a centrifugal microdroplet generation device and a digital immuno-PCR assay. Miniaturized centrifugal devices enable emulsification of hundreds of samples within 3 minutes using a common centrifuge. The bead-free digital immuno-PCR assay not only eliminates the need for multistep washing, but also possesses ultra-high detection sensitivity and accuracy. We characterized the performance of CDPro using recombinant interleukins (IL-3 and IL-6) as example targets and reported a LoD of 0.0128 pg mL−1. We also quantified IL-6 from 7 human clinical blood samples using the CDPro with only 0.5 μL plasma, which showed excellent agreement with an existing clinical protein diagnostic system with 25 μL plasma from those samples (R2 = 0.98).

Graphical abstract: Highly parallel, wash-free, and ultrasensitive centrifugal droplet digital protein detection in sub-microliter blood

Supplementary files

Article information

Article type
Paper
Submitted
10 Бер 2023
Accepted
17 Тра 2023
First published
17 Тра 2023

Lab Chip, 2023,23, 2758-2765

Highly parallel, wash-free, and ultrasensitive centrifugal droplet digital protein detection in sub-microliter blood

Z. Tang, F. Lv, D. E. Reynolds, S. Zhang, S. Zheng, J. Ko, Y. Chen and Y. Wang, Lab Chip, 2023, 23, 2758 DOI: 10.1039/D3LC00205E

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