Adsorption Medium, Method for Production Thereof, and Use Thereof for Purification of Biomolecules

Abstract

The present invention relates to an adsorption medium, especially a chromatography medium, to a method for the production thereof, and to the use of the adsorption medium according to the invention or of an adsorption medium produced according to the invention for the purification of biomolecules.

Claims

1. An adsorption medium, especially a chromatography medium comprising a chromatic matrix; polymeric space elements which have been bonded to the surface of the chromatography matrix; and Polymer chains containing chromatographically active centers, wherein the polymer chains have been bonded to the polymeric spacer elements.

2. The adsorption medium as claimed in claim 1 wherein the chromatography matrix comprising a material selected from the group consisting of natural or synthetic fibers, (polymer) membranes, porous, polymeric monolithic shaped bodies, polymer gels, films, nonwovens, wovens and inorganic materials.

3. The adsorption medium as claimed in claim 1, wherein the polymeric spacer elements are selected from the group consisting of polyamines, polyalcohols, polythiols, poly(meth)acrylates, poly(metho)acrylamides, poly-N-alkyl(meth)acrylamides and also copolymers consisting of two or more of the above polymers, and copolymers consisting of one or more of the above polymers and of polymers which do not bear nucleophilic fuctional groups.

4. The adsorption medium as claimed in claim 3, wherein the polymeric spacer elements are polyamines.

5. The adsorption medium as claimed in claim 1, wherein the polymeric spacer elements do not have epoxy groups.

6. The adsorption medium as claimed in claim 1, wherein the adsorption medium has a degree of grafting from 1% to 50%.

7. The adsorption medium as claimed in claim 1, wherein the functional group density of the chromatography matrix is from 1.5 to 30 nm.sup.?2.

8. The adsorption medium as claimed in claim 1, wherein the chromatographically active centers of the polymer chains are selected from the group consisting of anionic and cationic groups, hydrophobic groups, affinity ligands, metal chelates and reactive epoxide, aldehyde, azlactone, N-hydroxysuccinimide and/or carbodiimide groups.

9. A method for producing an adsorption medium as claimed in claim 1, comprising the steps: (a) providing a chromatography matrix; (b) immobilizing polymeric spacer elements on the surface of the chromatography matrix, and (c) immobilizing polymer chains containing chromatographically active centers on the polymeric spacer elements.

10. A method for producing an adsorption medium as claimed in claim 9, wherein step (c) comprises the steps: (c1) immobilizing a polymerization initiator on the polymeric spacer elements; (c2) carrying out a spacer element-initiated polymerization of a monomer to form immobilized polymer chains on the polymeric spacer elements; and (c3) optionally modifying the immobilized polymer chains to from chromatographically active centers on the polymeric chains.

11. The method for producing an adsorption medium as claimed in claim 10, wherein the spacer element-initiated polymerization step (c2) is carried out by means of an atom transfer radical polymerization (ATRP).

12. The method for producing an adsorption medium as claimed in claim 10, wherein use is made in step (c2) of polymerization solution comprising a portion of not more than 3 mol % of bifunctional monomers, based on the total amount of monomers in the solution.

13. The use of the adsorption medium as claimed in claim 1 for the purification of biomolecules.

14. The use as claimed in claim 13, wherein the biomolecules are proteins, peptides, amino acids, nucleic acids, viruses, virus-like particles and/or endotoxins.

Description

THE FIGURES SHOW

[0192] FIG. 1: Schematic breakthrough of a titration with 10 mM HCl solution.

[0193] FIG. 2: Calibration curve for determining the amount of bromide in a sample.

[0194] FIG. 3: Permeability as a function of chain length.

[0195] FIG. 4: Permeability as a function of initiator density.

[0196] FIG. 5: Permeability as a function of ligand density.

[0197] FIG. 6: Dependence of the ratio of static lysozyme-binding capacity to ionic capacity on the amine group density.

[0198] FIG. 7: Ratio of static protein-binding capacity to ionic capacity.

[0199] FIG. 8: Ratio of static protein-binding capacity to ionic capacity on a regenerated, crosslinked cellulose membrane.

[0200] FIG. 9: Ratio of static protein-binding capacity to ionic capacity on PBT Winged Fibers containing a polyallylamine spacer.