Abstract: This article presented a method,by which BSA and transferrin were simultane-ously separated and purified from adult bovine serum. To meet the requirements in serum-freecell culture,some quality indexes were examined,such as protein purity,endotoxin and fatty-acid contents,and compared with the products of Sigma Co.The experiments with cell cultureproved that BSA and transferrin gained by the method can successfully support animal cellgrowth when they are supplemented in serum-free medium.