3D printed microfluidic device for automated, pressure-driven, valve-injected microchip electrophoresis of preterm...

A 3D printed, automated, pressure-driven injection microfluidic system for microchip electrophoresis (µCE) of preterm birth (PTB)-related peptides and proteins has been developed. Functional microvalves were formed, either with a membrane thickness of 5 µm and a layer exposure time of 450 ms or with a membrane thickness of 10 µm and layer exposure times of 300-350 ms. These valves allowed for control of fluid flow in device microchannels during sample injection for µCE separation. Device design and µCE conditions using fluorescently labeled amino acids were optimized. A sample injection time of 0.5 s and a separation voltage of 450 V (460 V/cm) yielded the best separation efficiency and resolution. We demonstrated the first µCE separation with pressure-driven injection in a 3D printed microfluidic device using fluorescently labeled PTB biomarkers and 532 nm laser excitation. Detection limits for two PTB biomarkers, peptide 1 and peptide 2, for an injection time of 1.5 s were 400 pM and 15 nM, respectively, and the linear detection range for peptide 2 was 50-400 nM. This 3D printed microfluidic system holds promise for future integration of on-chip sample preparation processes with µCE, offering promising possibilities for PTB risk assessment.