A novel benzenediamine-benzodithiophene polymer is synthesized for use in biosensor fabrication for the detection of testosterone. The sensory platform is constructed via drop coating on a screen-printed carbon electrode, using poly(benzenediamine-Bis[(2-ethylhexyl)oxy]benzodithiophene) (pBDBT) as the polymer layer. Testosterone antibodies are immobilized on the polymer-coated electrode surface via glutaraldehyde, which binds to the surface through the amino functional groups on the polymer backbone. The changes in the surface features due to testosterone binding are investigated via electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry as well as contact angle measurements. Surface morphology of the modified electrodes is characterized by atomic force microscopy. The linear range and limit of detection of the sensor are calculated. Impact of possible interfering compounds is investigated. Furthermore, the sensory platform is utilized for testosterone analysis in synthetic biological fluids.