In this paper we demonstrate synthesis and characterization of MWCNTs embedded SU-8 electrospun nanofibers and their application towards ultrasensitive detection of cardiac biomarkers using Electrochemical Impedance spectroscopy (EIS). The composite nanofibers have excellent electrical and transduction properties owing to the presence of MWCNTs in addition to ease of functionalization and biocompatibility, which can be attributed to the presence of SU-8. Thus the synthesized nanofibers are ideal candidates for sensitive biosensor applications. As a proof concept, the detection of cardiac biomarkers, Myoglobin (Myo), cardiac Troponin I (cTn I) and Creatine Kinase MB (CK-MB) is demonstrated. The synthesized nanofibers were functionalized with the antibodies of the biomarkers and the detection was carried using Electrochemical Impedance Spectroscopy, an excellent technique for understanding the adsorption kinetics. A minimum detection limit of nano-gram/ml is demonstrated using this nanobiosensor platform. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim