<p> A dual-channel label-free electrochemical immunosensing platform is proposed based on a 3D-printed microfluidic architecture and Ti₃C₂-AgNPs composite material, achieving synergistic innovation in structural design and material engineering. Structurally, stereolithography (SLA) 3D printing constructs a spatially separated dual-chamber microfluidic array integrated with screen-printed electrodes. This design enables independent liquid transport and electrical signal isolation across different analytical channels, fundamentally suppressing signal crosstalk during multi-target detection. To achieve high sensitivity, the electrode surfaces were modified with a novel Ti₃C₂-AgNPs nanocomposite that significantly enhances charge transfer kinetics by preventing Ti₃C₂ nanosheet aggregation through modulation of AgNPs interlayer spacing. This integrated platform was validated through the simultaneous quantification of two critical prostate cancer (PCa) biomarkers, prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA). This immunosensor mitigates signal crosstalk whilst exhibiting superior analytical performance, with a linear range of 0.1–1,000 ng mL⁻¹, sensitivity of 0.0036 µA mL ng⁻¹ for PSA and 0.0024 µA mL ng⁻¹ for PSMA, and detection limits of 0.045 ng mL⁻¹ for PSA and 0.041 ng mL⁻¹&#xa0;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 PCa patients and healthy controls. Consequently, the proposed dual-channel label-free electrochemical immunosensors (EIs), based on Ti₃C₂-AgNPs nanocomposites, holds substantial promise for clinical diagnostic applications, with potential for expansion to the ultrasensitive detection of other disease-related biomarkers.</p> Graphical Abstract <p></p>