In this work, we present a fabrication process for microneedle sensors made of polylactic acid (PLA), which can be utilized for the electrochemical detection of various biomarkers in interstitial fluid. Microneedles were fabricated by the thermal compression molding of PLA into a laser machined polytetrafluoroethylene (PTFE) mold. Sensor fabrication was completed by forming working, counter, and reference electrodes on each sensor surface by Au sputtering through a stencil mask, followed by laser dicing to separate individual sensors from the substrate. The devised series of processes was designed to be suitable for mass production, where multiple microneedle sensors can be produced at once on a 4-inch wafer. The operational stability of the fabricated sensors was confirmed by linear sweep voltammetry and cyclic voltammetry at the range of working potentials of various biochemical molecules in interstitial fluid.