Abstract: A process for the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture in which the contact, washing and eluting phases are performed on a binding material made from a ligand, containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm, the binding material being so constructed that at least one biological or related substance is retained on the binding material during the contact and washing phases. In one preferred embodiment the ligand is a reactive dye, especially a triazinyl dye, the matrix is agarose or silica and the spacer arm is a substituted aminohexyl group. The chromatographic procedure is preferably performed at 100-3500 psi, at a flow rate of 0.5-2.0 ml/min. The choice of washing and eluting solutions depends on the material to be separated. However buffer solution to wash the column and a desorbing agent to elute the material are preferred.