ELUTION OF BIOMOLECULES FROM MULTI-MODAL RESINS USING MES AND MOPS AS MOBILE PHASE MODIFIERS

• US 20200131493A1
• Filed: 12/23/2019
• Published: 04/30/2020
• Est. Priority Date: 10/24/2012
• Status: Active Grant

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Godawat, Rahul, Cummings, Daniel, Warikoo, Veena
• Granted Patent: US 11,203,747 B2
• Time in Patent Office: Days
• CPC Class Codes: A61B 50/00 Containers, covers, furniture or holders specially adapted for surgical or diagnostic appliances or instruments, e.g. sterile covers drapes specially adapted for instruments A61B46/10
• CPC Class Codes: A61B 50/30 Containers specially adapted for packaging, protecting, dispensing, collecting or disposing of surgical or diagnostic appliances or instruments for wound clamps A61B17/10; for ligaturing clips A61B17/122; packages or dispensers for surgical gloves or finger-stalls A61B42/40
• CPC Class Codes: A61J 1/16 Holders for containers
• CPC Class Codes: B01D 15/3847 Multimodal interactions
• CPC Class Codes: B01D 15/426 Specific type of solvent
• CPC Class Codes: C07K 1/165 mixed-mode chromatography
• CPC Class Codes: C12N 9/2465 acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12Y 302/01022 Alpha-galactosidase (3.2.1.22)

Abstract

The present invention relates to methods for purifying or enriching a biomolecule using multimodal resins and an elution buffer containing a Good'"'"'s buffer.

Claims

1. A method for purifying or enriching a biomolecule, said method comprising the steps:

(a) providing an aqueous solution containing said biomolecule;

(b) contacting said aqueous solution with a multimodal resin;

(c) optionally washing the multimodal resin with an aqueous washing buffer;

(d) eluting the biomolecule from the multimodal resin by an aqueous buffer solution,

wherein said aqueous buffer solution comprises one or more of;

(i) at least 15 mM of a Good'"'"'s buffer;

(ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;

(iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and

(iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol; and

(e) collecting biomolecule containing fractions, thereby obtaining the biomolecule in purified or enriched form.

2. The method according to claim 1, wherein the aqueous buffer solution comprises 15 mM to 2.5 M Good'"'"'s buffer.

3. The method according to claim 1 or claim 2, wherein the Good'"'"'s buffer is MES, MOPS, or Tris.

4. The method according to claim 1 or claim 2, wherein the Good'"'"'s buffer is MES.

5. The method according to claim 1 or claim 2, wherein the Good'"'"'s buffer is MOPS.

6. The method according to any one of claims 1 to 5, wherein the biomolecule is a protein.

7. The method according to claim 6, wherein the protein is an enzyme.

8. The method according to claim 7, wherein the enzyme is a glycosidase.

9. The method according to claim 8, wherein the glycosidase is an alpha-galactosidase.

10. The method according to claim 9, wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta.

11. The method according to any one of claims 1 to 10, wherein the multimodal resin is selected from the group consisting of Capto™

Adhere, Capto™

MMC, Pall PPA HyperCel™

, Pall HEA HyperCel™

, Pall MEP HyperCel™

, and Eshmuno®

HCX.

12. A purified biomolecule obtained by the method according to any one of claims 1 to 11.

35. An article of manufacture comprising:

(a) a packaging material or container;

(b) an aqueous buffer solution comprising one or more of;

(i) at least 15 mM Good'"'"'s buffer;

(ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;

(iii) between 0 and 400 mM in total concentration of urea, guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and

(iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol;

 

and

(c) a data carrier containing instructions for carrying out a method according to any one of claims 1 to 11.

36. The article of manufacture according to claim 35, wherein the data carrier is a label present on the packaging material or container or a packaging insert.

37. The article of manufacture according to claim 35 or claim 36, wherein the data carrier comprises written information, visual information, or computer-readable information.

38. The article of manufacture according to any one of claims 35 to 37, wherein the data carrier further comprises one or more of the following information:

(i) instructions for use of aqueous buffer solution;

(ii) quality information such as information about the lot/batch number, the manufacturing or assembly site or the expiry or sell-by date of the aqueous buffer solution;

or information concerning the correct storage or handling of the article;

(iii) information concerning the composition of the aqueous buffer solution;

(iv) information concerning the interpretation of information obtained when performing methods according to any one of claims 1 to 11;

(v) a warning concerning possible misinterpretations or wrong results when applying unsuitable methods; and

(vi) a warning concerning possible misinterpretations or wrong results when using unsuitable reagent(s) and/or buffer(s).

39. The article of manufacture according to any one of claims 35 to 38, further comprising one or more of the following components:

(d) a chromatography medium;

(e) a chromatography column; and

(f) a reference biomolecule usable for calibrating a method for purifying or enriching a biomolecule.

40. The article of manufacture according to claim 39, wherein said chromatography column is prepacked with said chromatography medium.

41. The article of manufacture according to any one of claims 39 to 40, wherein said chromatography medium is a multimodal chromatography resin.

42. The article of manufacture according to claim 41, wherein the multimodal chromatography resin is selected from the group consisting of Capto™

Adhere, Capto™

MMC, Pall PPA HyperCel™

, Pall HEA HyperCel™

, Pall MEP HyperCel™

, and Eshmuno®

HCX.

43. The article of manufacture according to any one of claims 39 to 42, wherein the reference biomolecule is a protein or a nucleic acid.

44. The article of manufacture according to any one of claims 35 to 43, wherein the aqueous buffer solution comprises between 15 mM and 2.5 M Good'"'"'s buffer.

45. The article of manufacture according to any one of claims 35 to 44, wherein the at least one Good'"'"'s buffer is selected from the group consisting of MES, MOPS, and Tris.

46. The article of manufacture according to any one of claims 35 to 45, wherein the Good'"'"'s buffer is MES.

47. The article of manufacture according to any one of claims 35 to 45, wherein the Good'"'"'s buffer is MOPS.

13. A preparation comprising:

(a) a biomolecule; and

(b) an aqueous buffer solution comprising one or more of;

(i) at least 15 mM Good'"'"'s buffer;

(ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;

(iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and

(iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol.

14. The preparation according to claim 13, wherein the aqueous buffer solution comprises 15 mM and 2.5 M Good'"'"'s buffer.

15. The preparation according to claim 13 or 14, wherein the at least one Good'"'"'s buffer is selected from the group consisting of MES, MOPS, or Tris.

16. The preparation according to claim 13 or 14, wherein the Good'"'"'s buffer is MES.

17. The preparation according to claim 13 or 14, wherein the Good'"'"'s buffer is MOPS.

18. The preparation according to any one of claims 13 to 17, wherein the biomolecule is a protein.

19. The preparation according to claim 18, wherein the protein is an enzyme.

20. The preparation according to claim 19, wherein the enzyme is a glycosidase.

21. The preparation according to claim 20, wherein the glycosidase is an alpha-galactosidase.

22. The preparation according to claim 21, wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta.

23. A method for desorbing a biomolecule from a multimodal resin, comprising contacting a biomolecule bound to a multimodal resin with an aqueous buffer, wherein said aqueous buffer solution comprises one or more of:

(i) at least 15 mM Good'"'"'s buffer;

(ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;

(iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and

(iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol.

25. The method of claim 23 or claim 24, wherein the aqueous buffer solution comprises between 15 mM and 2.5 M Good'"'"'s buffer.

26. The method of any one of claims 23 to 25, wherein the Good'"'"'s buffer is selected from the group consisting of MES, MOPS, and Tris.

27. The method of any one of claims 23 to 25, wherein the Good'"'"'s buffer is MES.

28. The method of any one of claims 23 to 25, wherein the Good'"'"'s buffer is MOPS.

29. The method of any one of claims 23 to 28, wherein the biomolecule is a protein.

30. The method of claim 29, wherein the protein is an enzyme.

31. The method of claim 30, wherein the enzyme is a glycosidase.

32. The method of claim 31, wherein the glycosidase is an alpha-galactosidase.

33. The method of claim 32, wherein the alpha-galactosidase is selected from the group consisting of agalsidase alpha and agalsidase beta.

34. The method of any one of claims 23 to 33, wherein the multimodal resin is selected from the group consisting of Capto™

Adhere, Capto™

MMC, Pall PPA HyperCel™

, Pall HEA HyperCel™

, Pall MEP HyperCel™

, and Eshmuno®

HCX.

24. A method for increasing the recovery rate of a biomolecule adsorbed on a multimodal resin by contacting said multimodal resin onto which said biomolecule has been adsorbed with an aqueous buffer solution, wherein said aqueous buffer solution comprises one or more of:

(i) at least 15 mM Good'"'"'s buffer;

(ii) between 0 and 400 mM alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine;

(iii) between 0 and 400 mM guanidinium chloride, guanidinium thiocyanate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonium acetate, sodium chloride, sodium citrate, sodium caprylate, sodium phosphate, sodium iodide, potassium chloride, potassium citrate, potassium sulfate, potassium phosphate, potassium iodide, magnesium chloride, magnesium sulfate, magnesium phosphate, calcium chloride, calcium sulfate, calcium phosphate, or taurine; and

(iv) between 1% and 60% (v/v) of ethylene glycol or propylene glycol or a mixture of ethylene glycol and propylene glycol.