Bioleaching studies were conducted to evaluate the extraction of copper from combined flotation concentrate and smelter dust using two different dust-addition strategies, non-fractional (simultaneously with concentrate) and fractional (step-wise dust addition) in stirred tank reactors. The highest copper extraction obtained with the fractional dust addition was up to 94% while extraction using the non-fractional method leveled off at approximately 44%. Thus, an increased copper recovery of about 50% was achieved with the fractional dust-addition method. It seems that the main reason for increased copper recovery using fractional dust addition could be the control and prevention of chalcopyrite passivation (according to SEM/EDS, XRD, and quantitative mineralogy data). This leads to its improved dissolution.