Sensitive detection of ALP enzyme activity using a two-step DNA amplification biosensor

The biosensor demonstrates a remarkably low detection limit of 1.7 × 10⁻⁹ U L.

• A broad linear detection range from 0 to 0.2 U L was achieved.
• High specificity and sensitivity were observed in biomolecular detection.
• The biosensor effectively quantified alkaline phosphatase activity in cervical cancer cells and HeLa cell lysates, even at a 10-fold dilution.
• Precise detection at the single-cell level was enabled by the biosensor's sensitivity.
• This technology may provide a robust platform for cancer diagnosis and enzyme inhibitor screening.

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