(13) These methods are often coupled with total protein quantification via colorimetric assays such as microBCA and Bradford. (12) Alternatively, tunable elastomeric pore sensing analyzes individual particles via the electrical impedance they impart at an aperture. Here, the imaging of light scattered from particles moving under Brownian diffusion is used to determine the hydrodynamic size and concentration. Typically, exosomes are characterized via nanoparticle tracking analysis (NTA). Our proof-of-concept findings support the adoption of dual-mode acoustic analysis of exosomes, leveraging both frequency and dissipation monitoring for use in bioanalytical characterization. QCM-D sensors functionalized with anti-CD63 antibodies formed a direct immunoassay toward CD63-positive exosomes in 75% v/v serum, exhibiting a limit-of-detection of 2.9 × 10 8 and 1.4 × 10 8 exosome sized particles (ESPs)/mL for frequency and dissipation response, respectively, i.e., clinically relevant concentrations. Exosomes expressing the transmembrane protein CD63 were isolated by size-exclusion chromatography from cell culture media. Here, we investigate label-free immunosensing, using a quartz crystal microbalance with dissipation monitoring (QCM-D), to detect exosomes by exploiting their surface protein profile. The limitations and scarcity of current exosome characterization approaches have led to a growing demand for translational techniques, capable of determining their molecular composition and physical properties in physiological fluids. Exosomes are endocytic lipid-membrane bound bodies with the potential to be used as biomarkers in cancer and neurodegenerative disease.
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