Preparation and characterization of a macroporous agarose monolith as a stationary phase in IMAC chromatography

摘要

A macroporous monolith used as stationary phase for the separation of biomolecules by immobilized metal ion affinity chromatography (IMAC), based on D5 agarose (D5) chemically modified was proposed. The characterization of physical properties was studied. Pressure drop was 0.4 MPa, being a very low value compared to other similar chromatographic supports. The adsorption/desorption process was carried out using bovine serum albumin (BSA) at pH 7.4 as a target protein. The monolith was re-used for 20 adsorption/desorption cycles and it was possible to verify that the average percentage of adsorption in all cycles was 89.65%. It was also possible to apply a model in order to obtain the kinetic adsorption constant (k(a)), desorption constant (k(d)) and equilibrium constant (K-e) by the proposed system. These results indicate that this system is governed by the adsorption process.