A method of calibrating a reversible-binding sensor for detecting an analyte includes: (i) varying the temperature of a first calibration solution from a first temperature (T1) to a second temperature (T2) while the first calibration solution is in contact with a sensing region of the sensor; (ii) determining the sensor output for the first calibration solution as a function of temperature; (iii) varying the temperature of a second calibration solution from a third temperature (T3) to a fourth temperature (T4) while the second calibration solution is in contact with the sensing region, the second calibration solution having a concentration of analyte which is different from that of the first calibration solution; (iv) determining the sensor output for the second calibration solution as a function of temperature; and (v) using the determined sensor output from steps (ii) and (iv) to calibrate the sensor.