The presence of serpentine slime coating on the chalcopyrite surfaces severely affects the chalcopyrite floatability. In this study, three biodegradable polymers, namely polyaspartic acid (PASP), polyepoxy succinic acid (PESA), and hydrolyzed polymaleic (HPMA) were used as dispersants to disperse the chalcopyrite and serpentine particles. The micro-flotation experiments demonstrated that the application of dispersants in the flotation effectively reduced the negative impact of serpentine over a wide pH range, restoring the recovery rate of chalcopyrite to about 85%. The XPS analysis revealed that the dispersants could be adsorbed onto the mineral surfaces through the binding of carboxyl groups with the magnesium or copper sites. As a result, the surface potentials of serpentine changed from positive to negative, while that of the chalcopyrite became even more negative, resulting in a strong electrostatic repulsion between them, which effectively eliminated the covering of the serpentine on the chalcopyrite surface. The AFM force measurements indicate a weak interaction of the polycarboxylic acids dispersants with the surface of chalcopyrite. Consequently, the adsorption of dispersants on the chalcopyrite surface has an insignificant impact on the subsequent adsorption of collectors, which is further supported by the adhesion force measurements between the air bubble and mineral surfaces. Therefore, the application of polycarboxylic acid dispersants in the flotation significantly enhanced the floatability of chalcopyrite in the presence of the serpentine fines.