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Protoporphyrin IX-based all-solid-state choline (Ch) ion-selective electrodes (ISEs) were fabricated and characterized. Poly (3,4-ethylene dioxythiophene) doped with poly (styrene sulfonate) (PEDOT/PSS) functioning as an ion-to-electron transducer was electropolymerized on the gold wire (0.5 mm diameter). The conductive polymer was covered with a Ch selective membrane containing protoporphyrin IX as an ionophore, which exhibited a lower detection limit of 0.49 μM with the potentiometric method. The Ch sensor performed a wide linear range from 1 μM to 1 mM, a fast response time of less than 5 s, and a decent selectivity of common inorganic and organic ions in the human body. Characteristics such as pH and temperature stability, life span, reproducibility and repeatability were also investigated to be satisfied. With the background of artificial cerebrospinal fluid, the recovery rate in 10<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><msup><mrow></mrow><mrow><mo>−</mo><mn>5</mn></mrow></msup></semantics></math></inline-formula> M of Ch solution was measured by the standard addition method, revealing the potential for biological application.