An Integrated Electrochemical Platform Based on Ti 3 C 2 -Ag NPs and 3D- Printed Microfluidics for Simultaneous Detection of PSA and PSMA
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The multiplexed detection of tumor biomarkers represents a transformative strategy to improve diagnostic accuracy in oncology. A significant to the clinical translation of multiplexed label-free electrochemical immunosensors (EIs) is the issue of analytical reliability, which is frequently compromised by signal crosstalk and insufficient sensitivity. In this work, we designed a 3D-printed, reconfigurable microwell array through a combination of screen-printing, stereolithography (SLA) 3D printing, and microfluidic technologies for interference-free multiplexed detection. To achieve high sensitivity, the electrode surfaces were modified with a novel Ti 3 C 2 -Ag NPs nanocomposite that significantly enhances charge transfer kinetics by preventing Ti 3 C 2 nanosheet aggregation through modulation of Ag NPs interlayer spacing. This integrated platform was validated through the simultaneous quantification of two critical prostate cancer biomarkers, prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA). The immunosensor demonstrated both the complete elimination of signal cross-talk and excellent analytical performance, including a wide linear range (0.1–1,000 ng·mL⁻¹), low sensitivities (0.0036 µA·mL·ng − 1 for PSA and 0.0024 µA·mL·ng − 1 for PSMA), and low limits of detection (0.045 ng·mL⁻¹ for PSA and 0.041 ng·mL⁻¹ for PSMA). Furthermore, this device exhibited exceptional repeatability, stability, and specificity. Clinical validation using human serum samples exhibited strong concordance with clinical reference methods, enabling precise discrimination between prostate cancer patients and healthy controls. Consequently, the proposed dual-channel label-free EI, based on Ti 3 C 2 -Ag NPs nanocomposites, holds substantial promise for clinical diagnostic applications, with potential for expansion to the ultrasensitive detection of other disease-related biomarkers.
