L-Cysteine is a semi-essential and proteinogenic amino acid, with the formula HO2CCH (NH2) CH2SH. L-cysteine is mostly found in protein-rich foods. In this study, catalysts consisting of cobalt (Co) supported on carbon nanotubes (CNTs) were prepared using the sodium borohydride (NaBH4) reduction method. These catalysts were characterized by means of energy-dispersive x-ray (EDX) with scanning electron microscopy (SEM), temperature-programmed reduction (TPR), and temperature-programmed oxidation (TPO) techniques. Characterization results confirmed the successful preparation of the desired catalyst. After preparing the Co/CNT catalysts, a highly efficient and sensitive electrochemical sensor was developed using a glassy carbon electrode (GCE) modified with the Co/CNT catalysts. The electrochemical behavior of the bare GCE and Co/CNT-modified GCE electrodes was investigated using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) in 0.1 M phosphate buffer solution (PBS) + L-cysteine. Electrochemical results showed that the Co/CNT-modified GCE electrode exhibited high sensitivity and selectivity compared to the bare GCE, with sensitivity of 0.0046 µA/µM, limit of detection of 0.2 µM, and limit of quantification of 0.6 µM. Furthermore, this electrode showed higher sensitivity values than those reported in the literature. Further interference studies were performed using CV and EIS to investigate interfering species in the serum environment, such as D-glucose, uric acid, L-tyrosine, and L-tryptophan. In conclusion, the results suggest that the Co/CNT-modified GCE electrode is a promising catalyst for the sensitive detection of L-cysteine.