Rapid single-molecule digital detection of protein biomarkers for continuous monitoring of systemic immune disorders

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Abstract

Digital protein assays have great potential to advance immunodiagnostics because of their single-molecule sensitivity, high precision, and robust measurements. However, translating digital protein assays to acute clinical care has been challenging because it requires deployment of these assays with a rapid turnaround. Herein, we present a technology platform for ultrafast digital protein biomarker detection by using single-molecule counting of immune-complex formation events at an early, pre-equilibrium state. This method, which we term "pre-equilibrium digital enzyme-linked immunosorbent assay" (PEdELISA), can quantify a multiplexed panel of protein biomarkers in 10 µL of serum within an unprecedented assay incubation time of 15 to 300 seconds over a 104 dynamic range. PEdELISA allowed us to perform rapid monitoring of protein biomarkers in patients manifesting post-chimeric antigen receptor T-cell therapy cytokine release syndrome, with ∼30-minute sample-to-answer time and a sub-picograms per mL limit of detection. The rapid, sensitive, and low-input volume biomarker quantification enabled by PEdELISA is broadly applicable to timely monitoring of acute disease, potentially enabling more personalized treatment.

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APA

Song, Y., Sandford, E., Tian, Y., Yin, Q., Kozminski, A. G., Su, S. H., … Kurabayashi, K. (2021). Rapid single-molecule digital detection of protein biomarkers for continuous monitoring of systemic immune disorders. Blood, 137(12), 1591–1602. https://doi.org/10.1182/blood.2019004399

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