Abstract: A fully chromium loaded bovine low-molecular weight chromium-binding protein is isolated by a process that combines homogenization with supplementation of chromium content. Following homogenization with water, the homogenate is fractionated with ethanol, and the fractions obtained are subjected to serial chromatography (ion-exchange followed by size-exclusion chromatography) to obtain the biologically pure bovine LMWCr. This biologically pure material elutes from an HPLC column as essentially a single band, giving a high degree of purity. The LMWCr is useful as a dietary supplement, and for the treatment or prevention of a variety of chromium-related disease conditions.