The mechanism by which capsular polysaccharides inhibit phagocytosis is not clearly understood. We investigated the association between a negative surface charge and inhibition of phagocytosis by the capsular polysaccharide of Cryptococcus neoformans. A two-polymer aqueous-phase system containing phosphate ions was used to assess surface charge. Opsonins such as normal bovine serum and normal human immunoglobulin G reduced the surface charge on non-encapsulated cryptococci and simultaneously enhanced phagocytosis. These same opsonins had no effect on phagocytosis or surface charge of encapsulated cryptococci. F (ab')(2) fragments of normal human immunoglobulin G neither enhanced phagocytosis nor altered the surface charge of non-encapsulated cryptococci. Addition of purified cryptococcal polysaccharide to non-encapsulated cells inhibited phagocytosis of the yeast and induced a strong negative charge at the yeast surface. Chemical modification to reduce the surface charge of either purified cryptococcal polysaccharide or intact encapsulated cryptococci produced a small loss of phagocytosis-inhibiting activity; however, all treated polysaccharide preparations retained a significant ability to inhibit phagocytosis of the yeast. These results indicated that the association between surface charge and inhibition of phagocytosis was largely circumstantial, and presence of a negative surface charge could not account for the powerful antiphagocytic action of cryptococcal polysaccharide.