An electrochemical biosensor for sensitive detection of CKAP4 with application to the diagnosis of ovarian cancer

Abstract

Cytoskeletal remodeling is crucial in tumor progression and metastasis, with cytoskeleton-associated protein 4 (CKAP4) being a key protein involved this process; however, its detection remains a challenge. In this paper, we report a solid-state electrochemistry-enhanced biosensor based on Cu-TCPP nanosheets for sensitive detection of CKAP4. In the meantime, the proposed biosensor can present high specificity in CKAP4 recognition. Specifically, upon recognition of the target, the designed catalytic hairpin assembly (CHA) reaction is successfully initiated, generating lots of DNA duplexes, which can be cleaved by Exo III to release numerous truncated thiolated signal DNA (sDNA). Subsequently, the generated sDNA is introduced to a high-conductivity electrode surface modified with Cu-TCPP nanosheets which are loaded with methylene blue. Moreover, the introduction of sDNA triggers ligand displacement via competitive coordination on this surface, thereby forming non-electroactive Cu-sDNA complexes, which significantly modulates the current signals, enabling sensitive quantification of CKAP4 levels. The biosensor may achieve a low detection limit of 0.30 pg/mL, with a broad linear range covering five orders of magnitude, and exhibits satisfying anti-interference ability in complex biological matrices. Furthermore, it displays outstanding discriminatory accuracy in differentiating ovarian cancer patient samples from healthy controls, showing great potential for cancer diagnosis.