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Quantum relaxometry for detecting biomolecular interactions with single NV centers

MetadataDetails
Publication Date2025-08-25
JournalProceedings of the National Academy of Sciences
AuthorsMin Li, Qi Zhang, Xi Kong, Sheng Zhao, Baishen Pan
InstitutionsNanjing University, Hefei National Center for Physical Sciences at Nanoscale

The investigation of biomolecular interactions at the single-molecule level has emerged as a pivotal research area in life science, particularly through optical, mechanical, and electrochemical approaches. Spins existing widely in biological systems offer a unique degree of freedom for detecting such interactions. However, most previous studies have been largely confined to ensemble-level detection in the spin degree. Here, we developed a molecular interaction analysis method approaching single-molecule level based on relaxometry using the quantum sensor, nitrogen-vacancy (NV) center in diamond. Experiments utilized an optimized diamond surface functionalized with a polyethylenimine nanogel layer, achieving <mml:math xmlns:mml=ā€œhttp://www.w3.org/1998/Math/MathMLā€ display=ā€œinlineā€ overflow=ā€œscrollā€> <mml:mo>∼</mml:mo> </mml:math> 10 nm average protein distance and mitigating interfacial steric hindrance. Then we measured the strong interaction between streptavidin and spin-labeled biotin complexes, as well as the weak interaction between bovine serum albumin and biotin complexes, at both the micrometer scale and nanoscale. For the micrometer-scale measurements using ensemble NV centers, we reexamined the often-neglected fast relaxation component and proposed a relaxation rate evaluation method, substantially enhancing the measurement sensitivity. Furthermore, we achieved nanoscale detection approaching single-molecule level using single NV centers. This methodology holds promise for applications in molecular screening, identification, and kinetic studies at the single-molecule level, offering critical insights into molecular function and activity mechanisms.