A novel Bisphenol A (4,4'-isopropylidenediphenol, BPA) sensor was developed harnessing an electrochemical platform comprising a layer-by-layer assembled reduced graphene oxide (rGO) electrode and a designer probe specifically recognizing BPA. The BPA detection probe, a recombinant protein (LacI-BPA), was constructed by fusing a disulfide-constrained high affinity BPA binding peptide (CKSLENSYC) to the C-terminus of Lac repressor (LacI). Following expression and purification, the LacI-BPA was heat-denatured on-purpose to facilitate its direct adhesion on the rGO electrode surface via pi-stacking interaction. When the performance of the fabricated BPA sensor (LacI-BPA/rGO) was assessed by electrochemical impedance spectroscopy (EIS), it showed a wide linear dynamic range of BPA detection spanning from 100 fM to 10nM. Moreover, our BPA sensor exhibited negligible cross reactivity to BPA analogs such as Bisphenol S (BPS) and Bisphenol F (BPF) and almost complete spike recovery of BPA from plastic extracts containing various potential interferents. With these merits, the BPA sensor developed in the present study is expected to find practical application in selective and sensitive detection of BPA from diverse sample solutions.
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