Experimental data to reproduce the studies performed in the article "Disposable electrochemical aptasensor for rapid and selective vancomycindetection in clinical samples: Bridging affinity selection, computational modeling and clinical validation". It comprises information about the SELEX process (evaluation by UV-VIs spectrometry of the target immobilization efficiency, sequences obtained by cloning-sequencing, thermodynamic parameters of these sequences under the operational conditions, characterization of binding affinity by SPR) and optimization of the aptasensor (immobilization procedure, aptamer concentration, blocking agent immobilization time and concentration and target incubation time), analytical performance (calibration, reproducibility, stability, selectivity) and clinical validation in spiked and clinical serum samples and statistics.