PREPARATION OF CHROMATOGRAPHIC MATRICES BY FREE-RADICAL ADDITION LIGAND ATTACHMENT TO ALLYL GROUPS

Chromatographic matrices were produced by free radical addition reacti ons between immobilised allyl groups and suitable ligands. Efficient a ddition and high ligand densities were readily obtained with mercaptoa cids, glutathione, mercaptoethanol and sodium bisulphite, using aqueou s solvation. Addition of nitrogen containing thiol ligands other than glutathione was also demonstrated, although heat or radiation catalysi s was required. Addition of cysteamine or mercaptoacids resulted in sp acer arm derivatives, suitable for further ligand attachment chemistry . Mercaptoacetic acid and bisulphite attachment was used to prepare ca tion-exchange matrices. Glutathione derivatives were compared with mat rices prepared conventionally, for affinity chromatography of glutathi one transferase. (C) 1998 Elsevier Science B.V.