Development of an ATP-independent bioluminescent probe for detection of extracellular hydrogen peroxide

This work reports a new ATP-independent bioluminescent probe ( bor-DTZ ) for detecting hydrogen peroxide that is compatible with the Nanoluciferase enzyme. The probe is designed with an arylboronate ester protecting group appended to a diphenylterazine core via a self-immolative phenolate linker. Reaction with hydrogen peroxide reveals diphenylterazine, which can then react with Nanoluciferase to produce a detectable bioluminescent signal. Bor-DTZ shows a dose-dependent response to hydrogen peroxide and selectivity over other biologically relevant reactive oxygen species and can be applied to detect either intra- or extracellular species. We further demonstrate the ability of this platform to monitor fluxes in extracellular hydrogen peroxide in a breast cancer cell line in response to the anticancer treatment, cisplatin. The synthesis and evaluation of a new ATP-independent bioluminescent probe for H 2 O 2 detection is described and applied toward intra- and extracellular detection in a breast cancer cell model.