Media for separating small molecules from biomacromolecules in aqueous mixtures
A separation medium for removing small molecules from biomacromolecule in aqueous mixtures comprises gel filtration chromatography beads having a nominal protein fractional range of about 1000 Da to about 5000 Da and having an internal adsorbent matrix derived from a hydrophobicized scaffold. The gel filtration chromatography beads remove small molecules that are less than 1500 Da and have log Pow values greater than about −0.5 from biomacromolecules in aqueous mixtures. Devices containing the separation medium are also provided.
1. A separation medium comprising:
gel filtration chromatography beads having a nominal protein fractional range of about 1000 Da to about 5000 Da and having an internal hydrophobicized adsorbent matrix having attached aromatic or aliphatic moieties and that is derived from a hydrophilic polymeric scaffold that has been hydrophobicized, wherein the gel filtration chromatography beads remove small molecules that are less than 1500 Da and have log P values greater than about −0.5 from biomacromolecules in aqueous mixtures.
2. The invention of claim 1 , wherein the gel filtration chromatography beads are comprised of crosslinked polysaccharide.
3. The invention of claim 1 , wherein the gel filtration chromatography beads are comprised of polyacrylamide.
4. The invention of claim 1 , wherein the biomacromolecules include a protein.
5. The invention of claim 1 , wherein the biomacromolecules include an oligonucleotide.
6. The invention of claim 1 , wherein the biomacromolecules includes a nucleic acid duplex.
7. The invention of claim 1 , wherein the biomacromolecules include a hydrogel.
8. The invention of claim 1 , wherein the biomacromolecules include an antibody-drug conjugate.
9. The invention of claim 1 , wherein the scaffold is a polyacrylamide.
10. The invention of claim 1 , wherein the scaffold is a polyamide.
11. The invention of claim 1 , wherein the internal adsorbent matrix contains attached phenyl groups.
12. The invention of claim 1 , wherein the internal adsorbent matrix contains attached biphenyl groups.
13. The invention of claim 1 , wherein the internal adsorbent matrix contains attached naphthyl groups.
14. The invention of claim 1 , wherein the internal adsorbent matrix contains attached benzyl groups.
15. The invention of claim 1 , wherein the internal adsorbent matrix contains attached phenyl groups.
16. The invention of claim 1 , wherein the internal adsorbent matrix contains attached 9-fluorenylmethyl groups.
17. A separation medium comprising:
gel filtration chromatography beads having a nominal protein fractional range of about 1000 Da to about 5000 Da and having an internal hydrophobicized adsorbent matrix that is derived from a hydrophilic polymeric scaffold that has been hydrophobicized, wherein the gel filtration chromatography beads remove small molecules that are less than 1500 Da and have log P values greater than about −0.5 from biomacromolecules in aqueous mixtures; and
wherein the internal adsorbent matrix contains attached N-alpha-N-epsilon-bis-benzyloxycarbonyl-L-lysine groups.
18. The invention of claim 1 , wherein the internal adsorbent matrix contains attached N-benzyloxycarbonyl-L-leucine groups.
19. The invention of claim 1 , wherein the internal adsorbent matrix contains attached hexyl groups.
20. The invention of claim 1 , wherein the internal adsorbent matrix contains attached octyl groups.
21. A device for removing small molecules less than 1500 Da from aqueous protein solutions comprising:
a first layer of medium and a second layer of medium arranged in a column for a flow of fluid through the column with the first layer of medium disposed upstream from the second layer of medium in relation to the direction of fluid flow through the column;
the first layer of medium comprising gel filtration chromatography beads having a nominal protein fractional range of about 1000 Da to about 5000 Da and having an internal hydrophobicized adsorbent matrix having attached aromatic or aliphatic moieties that is derived from hydrophilic polymeric scaffold that has been hydrophobicized; and
the second layer of medium comprising a gel filtration chromatography medium;
wherein the gel filtration chromatography beads remove small molecules that are less than 1500 Da and have log P values greater than about −0.5 from biomacromolecules in aqueous mixtures.