In this study, an innovative flotation technology consisted of Cu differential flotation with high alkali lime and pyrite recovery with acid mine drainage (AMD) activation was investigated for the cleaner beneficiation of the copper sulfide ore. Flotation test results showed that H2SO4-CuSO4 and AMD could effectively activate the pyrite flotation with SBX collector. Moreover, the recovery of S concentrate is increased by 5.33% in the AMD system. Adsorption amount results of SBX collector indicated that the hydrophilic species (Ca2+, CaOH+ and FeOOH) were formed on the pyrite surfaces in the high alkali lime craft (pH=11.3) and degraded the interaction between SBX and pyrite surfaces. AMD can effectively clear off the hydrophilic calcium species and the copper ions originated from the AMD absorb onto the pyrite surfaces, facilitating the SBX collector adsorption. Composition analysis results of tailings water confirmed that the tailing water obtained by the AMD flotation system was more desirable to be recycled in the Cu differential flotation due to its higher pH value (8.7). The present study provides a novel approach for the treatment of AMD, and has the vital practical significance for the emission reduction of AMD and the increase of beneficiation profits.