AFFINITY CHROMATOGRAPHY MATRIX

• US 20160168229A1
• Filed: 07/04/2014
• Published: 06/16/2016
• Est. Priority Date: 07/05/2013
• Status: Active Grant

• Current Assignee: Laboratoire francais du Fractionnement et des Biotechnologies
• Original Assignee: Laboratoire francais du Fractionnement et des Biotechnologies
• Inventors: Paolantonacci, Philippe, Chtourou, Abdessatar
• Granted Patent: US 10,611,826 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 39/39525 Purification
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01D 15/3809 of the antigen-antibody type, e.g. protein A, G, L chromatography
• CPC Class Codes: B01J 20/265 modified or post-treated polymers polymer carriers or substrates subjected to further impregnating or coating B01J20/3208
• CPC Class Codes: B01J 20/289 bonded via a spacer
• CPC Class Codes: B01J 20/291 Gel sorbents
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3251 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 20/3274 Proteins, nucleic acids, polysaccharides, antibodies or antigens
• CPC Class Codes: B01J 2220/80 Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C07K 16/065 Purification, fragmentation
• CPC Class Codes: C07K 16/34 against blood group antigens
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

The invention relates to an affinity chromatography matrix, as a gel, comprising polymeric particles on which at least one oligosaccharide corresponding to a blood group A epitope and/or blood group B is grafted, via a spacer, characterized in that the density of oligosaccharides is comprised between 0.2 and 0.7 mg/ml of matrix. The invention also relates to the uses of this matrix for preparing concentrates of immunoglobulins for therapeutic use.

Claims

1. An affinity chromatography matrix, in form of a gel, comprising polymeric particles on which at least one oligosaccharide corresponding to a blood group A and/or group B epitope is grafted, said oligosaccharide being grafted to said particles via a spacer, wherein the density of oligosaccharides is comprised between 0.2 and 0.7 mg/ml of matrix and said spacer has a formula (I) —

NH—

R1-CO—

NH—

R2-, wherein R1 is a C₄-C₆ alkyl group, R2 is a C₃-C₈ alkyl group, and said spacer is bound through its amine function to the particle.

2. The matrix according to claim 1, comprising (i) polymeric particles on which at least one oligosaccharide corresponding to a blood group A epitope is grafted, and/or (ii) polymeric particles on which at least one oligosaccharide corresponding to a blood group B epitope is grafted.

3. The matrix according to claim 2, comprising (i) polymeric particles on which at least one oligosaccharide corresponding to a blood group A epitope is grafted, and (ii) polymeric particles on which at least one oligosaccharide corresponding to a blood group B epitope is grafted.

4. The matrix according to claim 1, comprising polymeric particles on which both at least one oligosaccharide corresponding to a blood group A epitope and at least one oligosaccharide corresponding to a blood group B epitope, are grafted.

5. The matrix according to claim 1, comprising a mixture of (i) polymeric particles on which at least one oligosaccharide corresponding to a blood group A epitope is grafted, (ii) polymeric particles on which at least one oligosaccharide corresponding to a blood group B epitope is grafted, and (iii) polymeric particles on which both at least one oligosaccharide corresponding to a blood group A epitope, and at least one oligosaccharide corresponding to a blood group B epitope, are grafted.

6. The matrix according to claim 1, wherein the density of oligosaccharides is comprised between 0.3 and 0.4 mg/ml of matrix.

7. The matrix according to claim 1, wherein the oligosaccharide corresponding to a blood group A epitope and/or the oligosaccharide corresponding to a blood group B epitope is a trisaccharide.

8. The matrix according to claim 7, wherein the oligosaccharide corresponding to a blood group A epitope is a trisaccharide N-acetylgalactosamine (GalNAc)-Galactose (Gal)-Fucose.

9. The matrix according to claim 7, wherein the oligosaccharide corresponding to a blood group B epitope is an oligosaccharide Galactose-Galactose-Fucose.

10. The matrix according to claim 1, wherein R1 is a C₅ alkyl group.

11. The matrix according to claim 1, wherein R2 is a C₃ alkyl group.

12. The matrix according to claim 3, wherein the polymeric particles on which at least one oligosaccharide corresponding to a blood group A epitope is grafted, and the polymeric particles on which at least one oligosaccharide corresponding to a blood group B epitope is grafted, are mixed in a proportion of 25/75 to 75/25 (v/v), preferably about 50/50 (v/v).

13. The matrix according to claim 1, wherein the polymer is a crosslinked polymer.

14. The matrix according to claim 13, wherein the polymer is cellulose.

15. A method of using the matrix as defined in claim 1, in an affinity chromatography binding anti-A and/or anti-B antibodies.

16. The method of using the matrix according to claim 15, for producing on an industrial scale immunoglobulins G (IgG) for therapeutic use.

17. A process for preparing a concentrate of immunoglobulins G (IgG) for therapeutic use, comprising the obtaining of an Ig composition from blood plasma, by ethanol fractionation and/or caprylic fractionation and/or chromatographic separation, and removal of the anti-A and/or anti-B antibodies possibly present in the composition, by means of an affinity chromatography applying the matrix as defined in claim 1.

18. The matrix according to claim 6, wherein the density of oligosaccharides is about 0.3 mg/ml of matrix.

19. The matrix according to claim 14, wherein the particles are porous cellulose beads.