Impedance Spectroscopy-Based Detection of Cardiac Biomarkers on Polyaniline Coated Filter Paper

Abstract

Cardiac biomarker detection helps in early diagnosis of cardiac syndromes. However, a rapid, cost-effective, reliable, and easy-to-use sensor is still unavailable. Herein, we report a disposable, label free, impedance-based biosensor for the detection of two common cardiac biomarkers, Myoglobin (Myo), and Myeloperoxidase (MPO). The sensor is fabricated on Whatman filter paper coated with a conducting polymer, polyaniline (PAni). The PAni-coated paper is functionalized with glutaraldehyde, a homo-bi functional crosslinker, prior to the covalent attachment of antibodies specific to each biomarker protein. Non-specific active regions of the sensor are blocked by treatment with bovine serum albumin. Conductive silver paste is used to define electrodes for AC impedance measurement on the PAni coated filter paper. Binding of the target protein (Myo or MPO) to the specific antibody changes the impedance between the electrodes; this is detected using AC impedance spectroscopy. The impedance is measured using a frequency response analyzer with an AC excitation voltage of 100 mV in the frequency range of 100 Hz-100 kHz. We can successfully detect Myo and MPO in buffer solution within the concentration range of 100 ng/mL-50 μg/mL. Detection limit obtained for these cardiac biomarkers spiked in human serum is 1 μg/mL. Further blocking using human serum improves the lower detection limit to 500 ng/mL. The developed biosensor uses inexpensive materials and fabrication techniques, detects cardiac biomarkers in clinically relevant concentrations rapidly (i.e., within 20 mins), and can be disposed in an environment friendly manner, thus making the sensor suitable for diagnostic applications.