In this study, a planar split-ring resonator (SRR)-based RF biosensor was developed for label-free detection of biomolecules such as the prostate cancer marker, prostate specific antigen (PSA), and cortisol stress hormone. The biosensor has a resonance-assisted transducer and is excited by a time-varying magnetic field component of a local high-impedance microstrip line. The resulting device exhibits an intrinsic S21 resonance with a quality-factor (or Q-factor) of 50. For the biomolecular interaction, anti-PSA and anti-cortisol were immobilized on the gold surface of the resonator by a protein-G mediated bioconjugation process and corresponding frequency shifts of Δf1p=30±2 MHz (for anti-PSA) and Δf1c=20±3 MHz (for anti-cortisol) were observed. The additional frequency shift of each PSA and cortisol antigen with a 100pg/ml concentration was about 5±1.5MHz and 3±1MHz, respectively. From the experimental results, we confirmed that our device is very effective RF biosensor with a limit of detection (LOD) of 100pg/ml and has sufficiently feasibility as a label-free biosensing scheme.