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

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...

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Veröffentlicht in:Blood 2021-03, Vol.137 (12), p.1591-1602
Hauptverfasser: Song, Yujing, Sandford, Erin, Tian, Yuzi, Yin, Qingtian, Kozminski, Andrew G., Su, Shiuan-Haur, Cai, Tao, Ye, Yuxuan, Chung, Meng Ting, Lindstrom, Ryan, Goicochea, Annika, Barabas, Jenny, Olesnavich, Mary, Rozwadowski, Michelle, Li, Yongqing, Alam, Hasan B., Singer, Benjamin H., Ghosh, Monalisa, Choi, Sung Won, Tewari, Muneesh, Kurabayashi, Katsuo
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Sprache:eng
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Zusammenfassung: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. •An innovative digital assay technology enables rapid, point-of-care measurement of multiple protein biomarkers in blood.•Continuous monitoring of protein blood biomarker profiles is shown for critically ill human patients with a digital assay. [Display omitted]
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.2019004399