Abstract: The present invention relates to a method of preparing an immobilised metal ion affinity chromatography (IMAC) adsorbent, which comprises to provide chromatography ligands comprised of alkylene diamine triacetic acid, or a derivative thereof, and coupling thereof to a carrier via nitrogen. In an advantageous embodiment, the alkylene diamine triacetic acid is ethylene diaminetriacetic acid (ED3A).

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.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of a) providing at least one cyclic scaffold comprising a carbonyl, an adjacent sulphur and a nucleophile; b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities are protected, on each scaffold by derivatisation of the nucleophile of said scaffolds, while retaining the cyclic structure of the scaffold; (c) ring-opening at the bond between the carbonyl and the sulphur of the derivatized scaffold by adding a reagent that adds one or more metal chelating affinity ligand arms to the scaffold; and, if required, (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment of the method, steps (c) and (d) are performed simultaneously as one single step.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of (a) providing at least one cyclic scaffold comprising a carbonyl, an adjacent sulphur and a nucleophile; (b) providing at least one polydentate metal chelating affinity ligand arm, optionally in a form wherein the metal chelating functionalities are protected, on each scaffold by derivatisation of the nucleophile of said scaffolds, while retaining the cyclic structure of the scaffold; (c) ring-opening at the bond between the carbonyl and the sulphur of the derivatised scaffold by adding a reagent that adds one or more metal chelating affinity ligand arms to the scaffold; and, if required, (d) deprotecting the functionalities of the ligand arm(s) provided in step (b). In the most preferred embodiment of the method, steps (c) and (d) are performed simultaneously as one single step.

Abstract: The present invention relates to a method of generating at least one polydentate metal chelating affinity ligand, which method comprises the steps of