Comparative study on aptamers as recognition elements for antibiotics in a label-free all-polymer biosensor.

We present an all-polymer electrochemical microfluidic biosensor using Topas(®) as substrate and a conductive polymer bilayer as electrode material. The conductive bilayer consists of tosylate doped poly(3,4-ethylenedioxythiophene) (PEDOT:TsO) and the hydroxymethyl derivative PEDOT-OH:TsO, which was covalently functionalized with two aptamer probes with affinity to ampicillin or kanamycin A, respectively. Using electrochemical impedance spectroscopy (EIS) we were able to detect ampicillin in a concentration range from 100pM to 1μM and kanamycin A from 10nM to 1mM. The obtained EIS spectra were fitted with an equivalent circuit model successfully explaining the impedance signal. Real samples from regular ultra-high temperature treated low-fat milk spiked with ampicillin were successfully tested to assess the functionality of the sensor with real samples. In conclusion, we have demonstrated the applicability of the newly developed platform for real time, label-free and selective impedimetric detection of commonly used antibiotics. Additionally it was possible to detect ampicillin in a milk sample at a concentration below the allowed maximum residue limit (MRL) in the European Union.