Understanding the interactions of proteins with humic acid in different media is essential for many applications. This paper investigates the interactions of bovine serum albumin with humic acid-Cu(II) complexes in a poly(hydroxyethylmethacrylate) cryogel column. Humic aggregates were immobilized within the cryogenic matrix. Protein sorption studies were conducted at different temperature, ionic strength, protein concentration, pH and flow-rate. Protein adsorption increased at lower pHs (close to the isoelectric point of the protein; 4.7) and higher temperatures but decreased at higher ionic strength and flow-rates. Both electrostatic and hydrophobic interactions played roles in the sorptive behavior of the protein molecules. Isotherm analysis showed the monolayer protein adsorption onto humic molecules immobilized column.