A compartmentalization-free microfluidic digital assay for detecting picogram levels of protein analytes

Abstract

Digitalizing the signals generated from single protein molecules has significantly improved the sensitivity of immunoassays compared to traditional analog “bulk” measurements. The single molecule array (Simoa) technology, for instance, leverages counting of single molecules on magnetic beads to detect low-abundance proteins in biofluids. While existing digital detection platforms are ultra-sensitive, they typically require compartmentalization and complex and bulky analysis equipment, limiting their applicability in resource-limited settings. Here, we introduce a compartmentalization-free digital detection technique, that allows for much more straightforward detection analysis. We applied this method to a model assay for detecting the SARS-CoV-2 spike protein and compared its performance to alternative techniques. We optimized the new method for digital microfluidics and present preliminary results using an automated system to analyze undiluted human saliva samples, with imaging performed on a portable optical system. We propose that future iterations of the scheme introduced here have the potential to enable a wide range of applications beyond the laboratory.

Graphical abstract: A compartmentalization-free microfluidic digital assay for detecting picogram levels of protein analytes

Supplementary files

Article information

Article type
Paper
Submitted
28 Jan 2025
Accepted
07 Apr 2025
First published
12 May 2025
This article is Open Access
Creative Commons BY license

Lab Chip, 2025, Advance Article

A compartmentalization-free microfluidic digital assay for detecting picogram levels of protein analytes

N. H. Le, N. Sathishkumar, A. Salari, R. Manning, R. E. Meyer, C. W. Kan, A. D. Wiener, M. A. Rossotti, S. Decombe, R. P. S. de Campos, M. D. Chamberlain, J. Tanha, N. R. Pollock, D. C. Duffy and A. R. Wheeler, Lab Chip, 2025, Advance Article , DOI: 10.1039/D5LC00103J

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