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Label-free aptasensors can be a promising point-of-care biosensor for detecting various cancer diseases due to their selectivity, sensitivity, and lower cost of production and operation. In this study, a highly sensitive aptasensor based on gold-covered polyethylene terephthalate electrodes (PET/Au) decorated with bipolar exfoliated graphene is proposed as a possible contender for disposable label-free aptasensor applications. Bipolar electrochemical exfoliation enables simultaneous exfoliation, reduction, and deposition of graphene nanosheets on prospective electrodes. Our comparative study confirms that the bipolar exfoliated graphene deposited on the negative feeding electrode (i.e., reduced graphene oxide) possesses better electrochemical properties for aptasensing. The optimized aptasensor based on bipolar exfoliated graphene deposited on PET/Au electrodes exhibits a highly sensitive response of 4.07 μA log c−1(unit ofc, pM) which is linear in the range of 0.0007-20 nM, and has a low limit of detection of 0.65 pM (S/N = 3). The aptasensor establishes highly selective performance with a stability of 91.2% after 6 days. This study demonstrates that bipolar electrochemistry is a simple yet efficient technique that could provide high-quality graphene for biosensing applications. Considering its simplicity and efficiency, the BPE technique promises the development of feasible and affordable lab-on-chip and point-of-care cancer diagnosis technologies.