A label free electrochemical detection method for the rapid detection of recombinant human erythropoietin (rhuEPO) has been developed. In this method, we modified the rhuEPO structure for its direct sensing without using a complex signal amplification strategy. The protein was selectively extracted from blood plasma sample using target-specific magnetic beads. After releasing rhuEPO from the magnetic beads, its disulfide bonds were electrochemically reduced and the protein was spontaneously assembled onto a nanostructured gold electrode via Au-S bonds formation. For electrochemical quantification, the reduced protein was desorbed from the electrode surface using differential pulse voltammetry (DPV). The desorption current was proportional to the concentration of rhuEPO in the range 1-1000 p.M. By cross-validating against ELISA, we found a 104.85 ± 3.35% agreement between the results obtained using the electrochemical biosensor and ELISA. Therefore the developed method has a strong potential for the sensitive detection of rhuEPO doping in sports as well as its rapid screening and pathology labs.
Keywords: Chronoamperometry; DPV; Disulfide bond reduction; Electrochemical biosensing; Nanostructured gold electrode; Recombinant human erythropoietin.
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