Selective binding of biological targets to solid phase ureides

Patent metadata

Field	Value
Publication number	US-9637724-B2
Application number	US-201414555288-A
Country	US
Kind code	B2
Filing date	Nov 26, 2014
Priority date	May 31, 2012
Publication date	May 2, 2017
Grant date	May 2, 2017

Abstract

A method of selectively separating a biological target from a sample including the biological target material or suspected of including the biological target includes the steps of (i) providing a solid including ureide moieties at its surface, (ii) contacting the sample with the solid, whereby a substantial fraction of the biological target in the sample binds to the ureide moieties, and (iii) separating the solid from the sample.

First claim

The invention claimed is:

1. A method of selectively separating a biological target from a sample comprising the biological target or suspected of comprising the biological target, the method comprising: (i) contacting the sample with a supersaturated allantoin, wherein at least 50% of the biological target in the sample binds to the supersaturated allantoin, thereby forming an undissolved precipitate comprising allantoin and the biological target; and (ii) separating the undissolved precipitate from the sample.

2. The method of claim 1 , wherein the biological target has a size in a range from about 10 nm to about 200 nm.

3. The method of claim 1 , wherein the biological target is a large biological target having a molecular weight in a range of about 10 megaDaltons or greater.

4. The method of claim 1 wherein the biological target comprises a cell.

5. The method of claim 4 , wherein the cell is a mammalian cell.

6. The method of claim 1 , wherein the biological target comprises a substructure of a cell.

7. The method of claim 6 , wherein the cellular substructure comprises an organelle.

8. The method of claim 6 , wherein the cellular substructure comprises an exosome.

9. The method of claim 1 , wherein at least 75% of the biological target binds to the supersaturated allantoin.

10. The method of claim 1 , wherein at least 90% of the biological target binds to the supersaturated allantoin.

11. The method of claim 1 , wherein at least 95% of the biological target binds to the supersaturated allantoin.

12. The method of claim 1 , wherein at least 99% of the biological target binds to the supersaturated allantoin.

13. The method of claim 1 , wherein substantially all of the biological target binds to the supersaturated allantoin.

14. The method of claim 1 , wherein the supersaturated allantoin comprises crystalline allantoin.

15. The method of claim 1 , wherein the supersaturated allantoin is in an aqueous liquid and comprises undissolved allantoin in the form of a solid by virtue of allantoin being present in the aqueous liquid at a supersaturating concentration.

16. The method of claim 1 , wherein the supersaturated allantoin is at a concentration greater than 0.56%.

17. The method of claim 1 , wherein the undissolved precipitate is separated from the sample by filtration or sedimentation.

18. The method of claim 17 , wherein, after (ii), the undissolved precipitate is washed by re-suspending the undissolved precipitate in a clean buffer then recovering the undissolved precipitate by sedimentation or filtration.

19. The method of claim 18 , wherein the clean buffer comprises allantoin at or close to saturation.

20. The method of claim 18 , wherein the washing step may be repeated one or more times.

21. The method of claim 19 , wherein the biological target is separated from at least some of the allantoin in the recovered undissolved precipitate by solubilizing the at least some of the allantoin.

22. The method of claim 21 , wherein the biological target is subsequently concentrated by diafiltration.

23. The method of claim 21 , wherein the solubilizing is performed simultaneous with concentrating the biological target by diafiltration.

24. The method of claim 22 , wherein the biological target is subsequently purified by an additional fractionation method.

25. The method of claim 1 , wherein the sample is a liquid or a gas.

26. The method of claim 1 , wherein the sample is a cell culture harvest containing cells or lysed cells.

27. The method of claim 26 , wherein the cells may have been previously removed.

28. The method of claim 1 , wherein the sample is aqueous.

29. The method of claim 1 , wherein the sample is water.

30. The method of claim 1 , wherein the sample is air.

31. The method of claim 1 , wherein the biological target is a desired biological target to be purified.

32. The method of claim 1 , wherein the biological target is a biological target to be removed from the sample.

33. The method of claim 32 , wherein the biological target may comprise more than one species of biological target to be removed from the sample.

34. The method of claim 32 , wherein the sample is treated with one or more species of soluble organic multivalent ions with the same net charge at an aggregate concentration ranging from 0.001 to 0.1%.

35. The method of claim 34 , wherein the one or more species of soluble organic multivalent ions comprises one or more species from the group of ethacridine, chlorhexidine, or octanoic acid at a concentration ranging from 0.001 to 0.1%.

36. The method of claim 32 , wherein the sample is treated with one or more species of immobilized organic multivalent ions of the same or different net charge, on a solid surface under conditions where the biological target does not bind to the immobilized organic multivalent ions.

37. The method of claim 32 , wherein the sample is treated with both soluble and immobilized organic multivalent ions, and wherein at least one species of immobilized organic multivalent ions has a net charge opposite to the soluble organic multivalent ions.

38. The method of claim 37 , wherein the immobilized organic multivalent ions include species of opposite net charge.

39. The method of claim 34 , wherein the one or more species of soluble organic multivalent ions comprises a metal affinity functionality.

40. The method of claim 37 , wherein treatment with the soluble organic multivalent ions occurs prior to treatment with the immobilized organic multivalent ions, or treatment with the soluble organic multivalent ions and treatment with the immobilized organic multivalent ions occurs essentially at the same time.

41. The method of claim 1 , wherein the method is practiced at a pH value of 4-10.

42. The method of claim 1 , wherein the method is practiced at a conductivity of 1-100 mS cm.

43. The method of claim 1 , wherein the sample is incubated together with the supersaturated allantoin for a period of time of 60 minutes.

44. The method of claim 1 , wherein the biological target is a virus being subjected to a treatment to inactivate the virus.

45. The method of claim 44 , wherein the virus inactivating treatment comprises exposure to a pH of 3 to 5.

46. The method of claim 1 wherein the biological target has a molecular weight of about 1 megaDalton to about 10 megaDaltons.

47. The method of claim 1 wherein the biological target is a protein.

48. The method of claim 1 , wherein the biological target is endotoxin.

49. The method of claim 1 wherein the biological target is a virus.

Assignees

• Agency Science Tech & Res

Inventors

• Gagnon Peter

Classifications

• C07K1/36

  by a combination of two or more processes of different types · CPC title

• B01J20/286

  Phases chemically bonded to a substrate, e.g. to silica or to polymers · CPC title

• C12N2770/24151

  Methods of production or purification of viral material · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• B01J20/3251

  comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur · CPC title