Method for preparation of a biomolecule adsorbent
View Patent ↗The present invention is relates to method for production of a biomolecule adsorbent with a pentadentate ligand with very strong chelating properties, and to methods for use thereof for, for example, protein purification, detection or binding. The biomolecule adsorbent comprises the formula: wherein Q is a carrier, S is a spacer, L is an amide linkage, X is COOH, and n=2 or 3.
1. A method of preparing a biomolecule adsorbent with a pentadentate ligand of the formula
wherein
Q is a carrier
S is a spacer
L is an amide linkage
X is COOH
n =2 or 3
comprising the following steps:
providing an alkylene diamine tetraacetic acid dianhydride;
providing an amino activated carrier;
coupling said alkylene diamine tetraacetic acid dianhydride to said carrier to form ligands comprised of alkylene diamine triacetic acid linked to said carrier via an amide linkage and a spacer; and
immobilizing charged metal ion ions by charging the adsorbent so obtained with metal ions, selected from Cu 2+ , Ni 2+ , Zn 2+ , Co 2+ , Fe 3+ and Ga 3+ to form pentadentate ligands.
2. The method of claim 1 , wherein Q comprises a cross-linked carbohydrate material, selected from agarose, agar, cellulose, dextran, chitosan, konjac, carrageenan, gellan, alginatec.
3. The method of claim 2 , in which Q is an amino-activated carbohydrate carrier.
4. The method of claim 3 , in which Q is an amino-activated agarose, and n is 2.
5. The method of claim 1 , wherein Q comprises cross-linked synthetic polymers, selected from styrene or styrene derivatives, divinylbenzene, acrylamides, acrylate esters, methacrylate esters, vinyl esters, vinyl amides.
6. The method of claim 1 , wherein the alkylene diamine tetraacetic acid dianhydride is EDTA dianhydride.
7. The method of claim 1 , wherein the spacer comprises at least 3 atoms.
8. The method of claim 1 , wherein Q comprises magnetic particles.