Separation matrix
The present invention relates to a separation matrix, which comprises a support; extenders coupled to an outer part of said support; and ligands coupled to said extenders, wherein the part of the support to which the extenders are coupled constitutes less than 50% of the volume of the separation matrix. The invention also embraces a method of preparing such a separation matrix, as well as a process wherein the separation media is used.
1. A separation matrix, comprising,
a porous support having an outer surface, an outermost part of the support, an interior part of the support, pores present in the outermost part, as well as pores present in the interior part of the support;
one or more extenders coupled to the outer surface as well as to the surfaces of the pores present in the outermost part of said support forming a shell portion of the support having a thickness in the range of 0.5-10 μm; and
two or more anion or cation exchange ligands coupled to each of said extenders,
wherein the molecular weight of the extenders is in the range of 50,000-500,000 Da and the extenders comprise multiple binding sites allowing for coupling of multiple ligands to each of the extenders;
wherein the shell portion of the support to which the extenders are coupled constitutes 1-17% of the total volume of the matrix, and
wherein there are substantially no ligands coupled to the pores of the interior part of the support.
2. The separation matrix of claim 1 , wherein the extenders are branched molecules.
3. The separation matrix of claim 1 , wherein the extenders are selected from the group consisting of agarose; dextran; dextrin; and a mixture of two or more thereof.
4. The separation matrix of claim 1 , wherein the support is comprised of substantially spherical particles.
5. The separation matrix of claim 1 , wherein the support is prepared from a polysaccharide.
6. The separation matrix of claim 1 , wherein the support is prepared from agarose.
7. The separation matrix of claim 1 , wherein the extender consists essentially of dextran.
8. The separation matrix of claim 1 , wherein the ligands are anion exchange ligands.
9. The separation matrix of claim 1 , wherein the ligands are cation exchange ligands.
10. The separation matrix of claim 1 , wherein the ligands are coupled to the extenders before the extenders are coupled to the support.
11. The separation matrix of claim 1 , wherein the separation matrix is spherical having a diameter of 10-500 μm.
12. A separation matrix, comprising,
a polysaccharide porous support having an outer surface, an outermost part of the support, an interior part of the support, pores present in the outermost part, as well as pores present in the interior part of the support;
one or more dextran extenders coupled to the outer surface as well as to the surfaces of pores present in the outermost part of said support forming a shell portion of the support having a thickness in the range of 0.5-10 μm; and
two or more anion or cation exchange ligands coupled to each of said dextran extenders,
wherein the molecular weight of the dextran extenders is in the range of 50,000-500,000 Da;
wherein the shell portion of the support to which the extenders are coupled constitutes 15-20% of the total volume of the matrix, and
wherein there are substantially no extenders coupled to the pores of the interior part of the polysaccharide support.
13. The separation matrix of claim 12 , wherein the extenders are branched dextrans.
14. The separation matrix of claim 12 , wherein the support is comprised of substantially spherical particles having a diameter of 100-200 μm.
15. The separation matrix of claim 12 , wherein the polysaccharide support is prepared from agarose.
16. The separation matrix of claim 12 , wherein the ligands are anion exchange ligands.
17. The separation matrix of claim 12 , wherein the ligands are cation exchange ligands.
18. The separation matrix of claim 12 , wherein the part of the support to which the extenders are coupled constitutes about 17% of the total volume of the matrix.
19. The separation matrix of claim 1 , wherein the dextran extenders are pre-allylated prior to support attachment or the ligands are coupled to the extenders before the extenders are coupled to the support.
20. A spherical separation matrix 100-200 μm in diameter, comprising,
an agarose porous support having an outer surface, an outermost part of the support, an interior part of the support, pores present in the outermost part, as well as pores present in the interior part of the support;
one or more dextran extenders coupled to the outer surface as well as to the surfaces of pores present in the outermost part of said support forming a shell portion of the support having a thickness in the range of 2-3 μm; and
two or more anion or cation exchange ligands coupled to each of said dextran extenders,
wherein the molecular weight of the dextran extenders is in the range of 50,000-500,000 Da;
wherein the shell portion of the support to which the extenders are coupled constitutes 15-20% of the total volume of the matrix, wherein the dextran extenders are pre-allylated prior to support attachment or the ligands are coupled to the extenders before the extenders are coupled to the support, and
wherein there are substantially no extenders coupled to the pores of the interior part of the agarose support.