Accurate and rapid biosensing is essential for patient diagnosis, pandemic response, and disease management; however, many biosensing techniques require complex preparation and processing steps. For example, enzyme-linked immunosorbent assay (ELISA) is considered the gold standard for quantitative biomarker detection, but the assay includes numerous blocking, labeling, and washing steps that require significant user engagement for large portions of the assay (up to several hours). We present a simple microparticle-based assay to rapidly isolate and purify biomarkers from whole blood in a handheld pipette for sensitive and streamlined biomarker detection. A key aspect of this technology is use of functionalized negative acoustic contrast particles (fNACPs), a unique set of acoustically responsive, modularly functionalized microparticles capable of capturing specific target biomarkers from whole blood. After capture, targets can be labeled in situ by fluorescent antibodies for subsequent analysis. The second major innovation is the handheld acoustic pipette that contains an acoustofluidic trapping channel; this channel establishes a half-wavelength acoustic standing wave by which fNACPs are efficiently separated from positive acoustic contrast species, including red and white blood cells. By integrating the fNACPs, pipette, and additional assay components, we demonstrate a rapid assay for detection of anti-OVA IgG from whole blood – with minimal sample preparation – all in under 75 minutes. The assay enables detection of anti-OVA at picomolar levels when analyzed by flow cytometry, a sensitivity that is competitive with commercial ELISAs.

Acoustically responsive microparticles in a handheld pipette for rapid and sensitive biosensing

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