Research for more reliable and cost-effective sensors in healthcare applications is a global priority. This is one of the main reasons that the scientific community is pushed in development of diagnostic tools for use in point-of-care (POC) applications. The combination of electrochemistry and 3D-printing techniques in terms of POC sensor development has many advantages. Utilization of ion selective electrodes (ISEs) in potentiometric techniques has provided fast and reliable analysis as well as high selectivity for analytes in many biofluids. 3D printing, as an additive manufacturing technique, has been used in a variety of applications since it has been shown to be a low-cost and rapid fabrication technique for many sensors that can be applied in diverse POC applications. As a result, 3D-printed electrochemical sensors are used for detection of a wide range of biomarkers associated with many diseases. Several 3D printed ISEs have already been shown to selectively detect analytes in biofluids, however a comprehensive study of the effects of biofluid fouling has yet to be established. Herein, we discuss the fabrication of a novel 3D printed sodium ISE to investigate the effect of biofouling on 3D printed ISEs in comparison to conventional PVC-based ISEs. Using a custom 3D printed microfluidic device, various biofluids were introduced to understand the effects of potential ISE deterioration after prolonged use in biofluids.