Thiacarbocyanine as ligand in dye-affinity chromatography for protein purification

A thiacarbocyanine has been post-grafted onto beaded cellulose by a curing method envisioning their use as a biomimetic ligand in dye-affinity chromatography. The immobilization of this dye was performed based on a previously brief derivatization study where the analogous microcrystalline cellulose was alternatively used and some of the curing reaction conditions were varied. The grafted beaded cellulose so obtained was qualitatively and quantitatively characterized by SEM, SEM-EDS and EA. The immobilized dye-affinity interaction with the standard proteins bovine serum albumin, alpha-chymotrypsin and lysosyme was analysed. The influence of the mobile phase composition on the chromatographic behaviour of these standard proteins was also studied. A selective interaction was observed allowing the separation of all the three proteins from an artificial mixture.