Affinity chromatography matrix

• US 10,611,826 B2
• Filed: 07/04/2014
• Issued: 04/07/2020
• 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
• Primary Examiner(s): Vasisth, Vishal V
• Application Number:
  US14/902,716
• Publication Number:
  US 20160168229A1
• Time in Patent Office: 2,104 Days
• Field of Search: 5303875, 252184, 536 179
• 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 via a spacer, wherein:

the density of oligosaccharides is from about 0.3 to about 0.4 mg/ml of the matrix and

said spacer has a formula provided by

NH—

R1-CO—

NH—

R2- 

 

(I),

wherein R1 is a C₅ alkyl group, R2 is a C₃ 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 oligosaccharide corresponding to a blood group A epitope and/or the oligosaccharide corresponding to a blood group B epitope is a trisaccharide.

7. The matrix according to claim 6, wherein the oligosaccharide corresponding to a blood group A epitope is a trisaccharide N-acetylgalactosamine (GalNAc)-Galactose (Gal)-Fucose.

8. The matrix according to claim 6, wherein the oligosaccharide corresponding to a blood group B epitope is an oligosaccharide Galactose-Galactose-Fucose.

9. 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).

10. The matrix according to claim 1, wherein the polymer is a cross-linked polymer.

11. The matrix according to claim 10, wherein the polymer is cellulose.

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

13. The method of using the matrix according to claim 12, for producing on an industrial scale immunoglobulins G (IgG) for therapeutic use.

14. A process for preparing a concentrate of immunoglobulins G (IgG) for therapeutic use, comprising obtaining an Ig composition from blood plasma, by ethanol fractionation and/or caprylic fractionation and/or chromatographic separation, and subjecting the Ig composition to affinity chromatography using the matrix of claim 1 to remove the anti-A and/or anti-B antibodies from the Ig composition.

15. The matrix according to claim 1, wherein the density of oligosaccharides is about 0.3 mg/ml of matrix.

16. The matrix according to claim 11, wherein the particles are porous, cross-linked cellulose beads.

17. 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 via a spacer, wherein:

the at least one oligosaccharide is a trisaccharide;

the density of oligosaccharides is from about 0.3 to about 0.4 mg/ml of the matrix;

said spacer has a formula provided by

NH—

R1-CO—

NH—

R2- 

 

(I),

wherein R1 is a C₅ alkyl group, R2 is a C₃ alkyl group, and said spacer is bound through its amine function to the particle; and

the particles are porous cross-linked cellulose beads.