Phosphorothioated oligonucleotides on gold-coated electrospun polymeric fibers for electrochemical genosensors

This study investigates the development of electrochemical genosensors using gold-coated electrospun polymeric fibers electrodes, Au/PMMA/PET and immobilized phosphorothioated oligonucleotides. Scanning electron microscopy (SEM) with energy-dispersive X-rays spectroscopy (EDS) revealed a uniform distribution of oligonucleotides on the fibers, contrary to planar gold electrodes Au/Ti/SiO2/Si, where network-like films were observed. X-ray photoelectron spectroscopy (XPS) confirmed the successful immobilization of the phosphorothioated oligonucleotides via strong covalent gold-sulfur bonds, while surface plasmon resonance (SPR) indicated superior binding affinity, with significantly lower equilibrium dissociation constants, when compared to unmodified probes. The detection of BCR/ABL fusion gene of chronic myeloid leukemia using differential pulse voltammetry and methylene blue as electroactive indicator, showed that the Au/PMMA/PET electrodes achieved a sensitivity of 379 +/- 12 mu A cm(-)(2) pM(-)(1) and a limit of detection of similar to 5.00 +/- 0.01 fM, outperforming the Au/Ti/SiO2/Si planar electrodes. Reduced non-specific adsorption was observed on the Au/PMMA/PET electrodes and attributed to the inherent charges introduced during the electrospinning process, which created localized electrostatic fields that repelled weakly adsorbing molecules. These findings demonstrate the potential of Au/PMMA/PET electrodes as a robust platform for further development of high-performance clinical diagnostic devices.