Agarose Derivatives for Purification of Protein by Affinity Chromatography

Abstract

AFFINITY chromatography^1–5, which in principle is related to the use of immunoadsorbants for the purification of antibodies⁶, exploits the reversible and specific interactions of ligands with macromolecules. Purification is affected by chromatographing the protein to be purified on a column containing an insoluble matrix to which the specific ligand is covalently attached. Beaded agarose (‘Sepharose’) is a useful insoluble support which has recently been used successfully in the selective purification of enzymes^1–5,7,9, antibodies and antigens^10–15, chemically synthesized peptides^16,17, thyroxine binding serum proteins¹⁸, and in the study of the interaction of insulin with cell membranes¹⁹. In these studies the ligands or peptides were attached through their amino groups after activation of ‘Sepharose’ with cyanogen bromide^20,21.