Double-layer vs single-layer cryogels as a dye affinity chromatography column, comparison study

Sammanfattning: Dye affinity chromatography is one of the chromatography techniques which uses dye as a ligand for protein purification. In this study, single-layer poly(acrylamide-allyl glycidyl ether) cryogel was prepared, which contains free epoxy groups at sub-zero temperature. The double-layer was prepared inside a primary single-layer cryogel to form a secondary network inside the gel. Cibacron Blue F3GA was immobilized onto single and double-layer cryogels by covalently binding with the epoxy groups. Bovine serum albumin (BSA) adsorption on the formed dye affinity single and double-layer cryogels was investigated. It was found that the kinetic model and adsorption isotherms follow the Pseudo-second-order and Freundlich model, respectively. The data showed that the binding capacity of double-layer cryogel decreased compared to single-layer cryogel due to lower surface area. However, the mechanical stability of the double-layer cryogel (based on Young's modulus) is higher than that of the corresponding single-layer cryogel. Both single and double-layer cryogels could be reused more than ten times in adsorption/elution cycles without losing their binding capacity. The physical and chemical properties of the cryogels were characterized by scanning electron microscopy, confocal microscopy, krypton adsorption, fourier transform infrared (FT-IR) spectroscopy, and texture analysis.