Hemin coupled protein biomimetic nanozymes combining confinement effects for sensitive chemiluminescence immunoassay

Abstract

Nanozymes offer superior stability and resistance to deactivation while mimicking the catalytic activity of natural enzymes, thereby expanding their applications across various fields. However, the development of nanozymes with both high catalytic activity and good biocompatibility remains a significant challenge. In particular, biocompatible nanozymes are highly desirable for immunoassays and biological detection. In this work, a series of nanozymes were prepared using bovine serum albumin (BSA) as a template and hemin as the catalytic active component. By activating the carboxyl groups of hemin, we compared the catalytic activities of nanozymes prepared by directly adding native BSA with those prepared using dBSA, obtained after spin-induced denaturation of BSA. Encapsulation of hemin by dBSA not only alleviated hemin aggregation and inactivation in aqueous solution but also enhanced its peroxidase-like activity through a confinement effect. As a result, the dBSA-hemin nanozymes efficiently catalyzed the luminol–H₂O₂ system, producing stable glow-type chemiluminescence. Taking advantage of the excellent catalytic activity of these nanozymes, a chemiluminescence immunosensor was developed using the nanozymes as signal labels for the detection of CA15–3. The constructed immunosensor exhibited a wide linear response range from 0.001 to 100 U/mL and a low detection limit of 0.0003 U/mL at a signal-to-noise ratio of 3. These results demonstrate that the use of simple materials and preparation methods remains highly promising. Moreover, the integration of self-assembly with confinement effects provides a new strategy for designing efficient nanozyme systems.

Keywords

Nanozymes / Hemin / Bovine serum albumin / CA15–3 / Chemiluminescence immunoassay

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

CRediT authorship contribution statement

Tong Jiang: Writing – original draft, Methodology, Investigation, Conceptualization. Yanan Lou: Writing – original draft, Data curation. Jin Wang: Investigation. Huan Wang: Methodology. Yuyang Li: Resources. Hongmin Ma: Writing – review & editing, Project administration, Funding acquisition.

Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (22374059), Ma thanks the Young Taishan Scholars Program of Shandong Province of China.

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