This paper presents a promising avenue of Radio Frequency (RF) biosensors for sensitive and real-time monitoring of creatinine detection. Knowing creatinine levels in the human body is related to its possible association with renal, muscular, and thyroid dysfunction. The detection was performed using an Inter-Digitated Capacitor (IDC) made of copper (Cu) metal over an FR4 substrate. To demonstrate our methodology, we have chosen Phosphate Buffer (PB) as our solvent for making the creatinine solutions of different concentrations. Moreover, Assayed Chemistry Control (ACC), a reference control consisting of human serum-based solutions has been mixed with the different concentrations of creatinine in a ratio of 1:9 to spike the creatinine value in the ACC solution. The sensor has been designed using a High-Frequency Structure Simulator (HFSS) tool with an operating frequency of 2.53 GHz. Then the design is fabricated over the FR4 printed circuit board (PCB) and tested using a Vector Network Analyzer (VNA). However, the sensitive area of the IDC is introduced to grade 4 Whatman filter paper for the Sample Under Test (SUT) handling unit. The main advantage of using Whatman filter paper is that the uniform spreading of liquid reduces experimental error, and less volume is required for testing the sample. The principal idea implemented in the biosensor design is to track the shift in the operating frequency in the presence of different concentrations of creatinine mix in ACC solution with Phosphate Buffer (PB) solution as a reference.