Abstract
It is necessary to develop reliable chemiluminescence strategies for determination of intracellular adenosine triphosphate (ATP), which is vital in life science and clinical diagnosis. However, the current chemiluminescence methods based on firefly luciferase suffered from low delivery efficiency, unsatisfied targeting performance, and autohydrolysis in living biosystem. To circumvent these drawbacks, a thermoresponsive polymer nanocomposite modified with firefly luciferase and ATP aptamer (PFLNC@aptamer) was fabricated, which targeted ATP and determined the intracellular ATP levels via measuring the chemiluminescence signals at different temperatures. The PFLNC@aptamer exhibited capability for the enzymolysis efficiency regulation, increased 21.0% with temperature change from 37.0 to 25.0 °C. The ATP detection limit was 3.3 nM with a linear relationship from 10.0 nM to 0.1 mM. Moreover, the thermoresponsive nanocomposite could also effectively avoid the interference during delivering firefly luciferase into the living cells and effectively discriminate ATP via the immobilized ATP aptamer, which further confirmed its reliability for practical applications. It paves a specific avenue for effective intracellular ATP monitoring in fundamental and applied research.
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