Abstract: The present invention relates to a method of preparing a separation matrix comprising at least one insoluble carrier to which sulphate ligands have been attached via extenders, which method comprises coupling, in a first step, of the sulphate ligands to the extenders and, in a subsequent step, attaching the extenders to an insoluble carrier. The invention also relates to a separation matrix comprised of at least one insoluble carrier to which sulphate ligands have been attached via extenders. Advantageously, no sulphate ligands are directly attached to the insoluble carrier. The extenders may be natural polymers, such as dextran, and the insoluble carrier may be made from natural polymers, such as agarose, or synthetic polymers. The invention also relates to a method of purifying virus, such as influenza virus, using a separation matrix according to the invention.

Abstract: The present invention relates to a method for generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of (a) providing at least one scaffold defined by the general formula (I): H2N—(X1)n—S—S—(X2)m—CH2—NH2 wherein X1 and X2 irrespective of each other are carbon atoms or heteroatoms, and n and m irrespective of each other are integers of 1 to 5; (b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities of at least one arm are protected, on each scaffold by derivatisation of the nucleophilic NH2 groups of the scaffold; (c) reducing the disulfide bond of the derivatised scaffold; and, if required (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment, the reduction of the disulfide bond and the deprotection step is performed essentially simultaneously.

Abstract: The present invention relates to a method of preparing a separation matrix comprising at least one insoluble carrier to which sulphate ligands have been attached via extenders, which method comprises coupling, in a first step, of the sulphate ligands to the extenders and, in a subsequent step, attaching the extenders to an insoluble carrier. The invention also relates to a separation matrix comprised of at least one insoluble carrier to which sulphate ligands have been attached via extenders. Advantageously, no sulphate ligands are directly attached to the insoluble carrier. The extenders may be natural polymers, such as dextran, and the insoluble carrier may be made from natural polymers, such as agarose, or synthetic polymers. The invention also relates to a method of purifying virus, such as influenza virus, using a separation matrix according to the invention.

Abstract: The present invention relates to a method for generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of (a) providing at least one scaffold defined by the general formula (I): H2N—(X1)n—S—S—(X2)m—CH2—NH2 wherein X1 and X2 irrespective of each other are carbon atoms or heteroatoms, and n and m irrespective of each other are integers of 1 to 5; (b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities of at least one arm are protected, on each scaffold by derivatisation of the nucleophilic NH2 groups of the scaffold; (c) reducing the disulfide bond of the derivatised scaffold; and, if required (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment, the reduction of the disulfide bond and the deprotection step is performed essentially simultaneously.

Abstract: The present invention relates to the preparation of a metal chelating separation medium, which comprises providing a compound defined by formula: R—O—CO—(CH2)n—CHNH2—CO—OH wherein R is a selectively cleavable group; and n is an integer of 1–4; reacting it with ethyl alcohol; derivatisation; cleavage of R; immobilisation of the resulting compound to a base matrix; and deprotection of the carboxylic acid. In one embodiment, the compound provided in the first step is ethyl ester of aspartic acid or of glutamic acid, and the derivatisation agent is bromo-acetic acid ethyl acetate.

Abstract: The present invention relates to the preparation of a metal chelating separation medium, which comprises providing a compound defined by formula: R—O—CO—(CH2)n—CHNH2—CO—OH wherein R is a selectively cleavable group; and n is an integer of 1-4; reacting it with ethyl alcohol; derivatisation; cleavage of R; immobilisation of the resulting compound to a base matrix; and deprotection of the carboxylic acid. In one embodiment, the compound provided in the first step is ethyl ester of aspartic acid or of glutamic acid, and the derivatisation agent is bromo-acetic acid ethyl acetate.