A major difficulty in the purification of many serum proteins is their separation from albumin. Human, rat and mouse alpha-fetoproteins (AFP) are physicochemically similar to the albumins of the respective species so that separation by conventional physical and chemical techniques is difticultL. Travis et dz described the removz&of albumin from human serum by &i&y chromatography on a column of agarose_ Cibacron Blue conjugate. We attempted to apply their method as a step'in the purification of non-human AFP and found a far lower &ii&y of rat, mouse and bovine albumin for Cibacron Blue than of human albumin under the same conditions. This paper describes conditions for chromatography of murine and bovine albumin on Cibacron Blue-agarose.
EXPERIMEN-TAL
-Cibacron Blue-agarosel was packed in a 240 x 9 mm coiuinn (bed vokme, 15 ml) and equilibrated with one of the following buffers: buffer I, 10 m.M Tris-KCE, pH 7.5; buffer 2,50 mM Tris-HCl, pH 7.5, EQ mM NaCl; brrffer 3,50 m&Z Tris-HCI;: pH 7.5,50 mM NaCl; buffer 4,50 mM Tris-HCl, phi 7.5,500 mM NaCI. All buffers contained 10 mM NaN,. Buffers 3 and 4 have been used to prepare albumin-free human serum proteins2*3.