We report on the lactate biosensor with linear calibration range from 0.5 to 100 mM, which encircles possible levels of this metabolite concentration in both human sweat and blood. The linear calibration range at high analyte concentrations, which exceeds the Michaelis constant of lactate oxidase by several orders of magnitude, is provided by an additional perfluorosulfonated ionomer diffusion membrane. In contrast to the known lactate biosensors, which retain their response within less than a couple of hours, the reported system displays 100% response for dozens of hours even upon high analyte concentrations. The biosensors with an additional diffusion-limiting membrane have been validated for lactate detection both in human blood serum and in undiluted human sweat shortly after its secretion. Both linear response in the entire range of blood and sweat lactate concentrations and ultrahigh operational stability would provide the use of the elaborated biosensor in wearable devices for the monitoring of hypoxia.