Space-borne differential optical absorption spectrometer is used for remote sensing of atmospheric trace gas global distribution. This instrument acquires high accuracy UV/Vis radiation scattered or reflected by air or earth surface, and can monitor distribution and variation of trace gases based on differential optical absorption spectrum algorithm. Spectral calibration is the premise and base of quantification of remote sensing data of the instrument, and the precision of calibration directly decides the level of development and application of the instrument. Considering the characteristic of large field, wide wavelength range, high spatial and spectral resolution of the space-borne differential optical absorption spectrometer, a spectral calibration method is presented, a calibration device was built, the equation of spectral calibration was calculated through peak searching and regression analysis, and finally the full field spectral calibration of the instrument was realized. The precision of spectral calibration was verified with Fraunhofer lines of solar light.