Nanoparticles for manipulation of biopolymers and methods of thereof

• US 20060177855A1
• Filed: 01/23/2006
• Published: 08/10/2006
• Est. Priority Date: 01/21/2005
• Status: Active Grant

• Current Assignee: Argela Technologies (Levent Yapilandirma Yönetimi Telekomünikasyon AS)
• Original Assignee: Argela Technologies (Levent Yapilandirma Yönetimi Telekomünikasyon AS)
• Inventors: Utermohlen, Joseph G., Hogan, Michael E., Diggins, Paul E.
• Granted Patent: US 7,964,380 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01D 15/3828 Ligand exchange chromatography, e.g. complexation, chelation or metal interaction chromatography
• CPC Class Codes: B01J 20/289 bonded via a spacer
• CPC Class Codes: B01J 20/3204 Inorganic carriers, supports or substrates
• CPC Class Codes: B01J 20/3272 Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3274 Proteins, nucleic acids, polysaccharides, antibodies or antigens
• CPC Class Codes: B01J 20/3289 Coatings involving more than one layer of same or different nature
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/54346 Nanoparticles

Abstract

Matrices for manipulation of biopolymers, including the separation, purification, bilization and archival storage of biopolymers is disclosed.

Claims

1. Solid-phase, non-porous particles for manipulating of biomolecules, comprising a surface area to volume ratio (m²/cm³) greater than 10, and a density (ρ

) greater than 2, and having sedimentation rates in water, with V_min greater than 0.1 cm/min at 10,000 G and V_max less than 2 cm/min at 500 G at standard temperature and pressures.

2. The particles of claim 1, said particles having a density greater than about 2 less than or equal to about 2.5, and effective spherical diameters as determined by two times Stokes radius in the range of 60 nm to 1000 nm.

3. The particles of claim 1, further having a density greater than about 2.5 less than or equal to about 3 and effective spherical diameters as determined by two times Stokes radius in the range of 40 nm to 800 nm.

4. The particles of claim 1, further having a density greater between about 3 to less than or equal to about 3.5 and effective spherical diameters as determined by two times Stokes radius between about 35 nm to 400 nm.

5. The particles of claim 1, further having a density greater than about 3.5 to less than or equal to about 4, and effective spherical diameters as determined by two times Stokes radius between about 20 nm to about 700 nm.

6. The particles of claim 1, further having a density greater than about 4 to less than or equal to about 4.5, and effective spherical diameters as determined by two times Stokes radius between about 30 nm to about 600 nm.

7. The particles of claim 1, further having a density greater than about 4.5 to less than or equal to about 5, and effective spherical diameters as determined by two times Stokes radius between about 25 nm to about 550 nm.

8. The particles of claim 1, further having a density greater than about 5 to less than or equal to about 5.5, and effective spherical diameters as determined by two times Stokes radius between about 25 nm to about 500 nm.

9. The particles of claim 1, further having a density greater than about 5.5 to less than or equal to about 6, and effective spherical diameters as determined by two times Stokes radius between about 25 nm to about 450 nm.

10. The particles of claim 1, further having a density greater than about 6 to less than or equal to about 6.5, and effective spherical diameters as determined by two times Stokes radius between about 20 nm to about 450 nm.

11. The particles of claim 1, further having a density greater than about 6.5 to less than or equal to about 7, and effective spherical diameters as determined by two times Stokes radius between about 20 nm to about 400 nm.

12. The particles of claim 1, further having a density greater than about 7 to less than or equal to about 7.5, and effective spherical diameters as determined by two times Stokes radius between about 20 nm to about 400 nm.

13. The particles of claim 1, further having a density greater than about 7.5 to less than or equal to about 14, and effective spherical diameters as determined by two times Stokes radius between about 15 nm to about 300 nm.

14. The particles of claim 1, further having a density greater than about 14 to less than or equal to about 20, and effective spherical diameters as determined by two times Stokes radius between about 12 nm to about 240 nm.

47. A kit comprising immunoglobulins isolated using the solid-phase particles according to claim 1.

48. A reagent for analysis and isolation comprising the solid-phase particles according to claim 1.

49. The solid-phase particles according to claim 1, wherein the metals are selected from the group consisting of Al, Ca, Co, Cu, Fe, Ga, Mg, Mn, In, Ni, Ti, and Zn.

50. A method of obtaining of proteins or nucleic acids comprising using the solid-phase particles according to claim 1.

59. The solid-phase particles according to claim 9, wherein the particles are kaolin particles.

15. Solid-phase particles comprising nanoparticles activated by a biopolymer binding agent for manipulating of biopolymers, wherein the solid phase particles are non-porous and comprise metals or semi-metals in the form of crystalline, amorphous, polymers or mixtures thereof, in colloidal solutions.

16. The solid-phase particles according to claim 15, wherein said solid phase particles have a surface area to volume ratio (m2/cm3) greater than 10, and a density (ρ

) greater than 2, and have sedimentation rates in water, with V_min greater than 0.1 cm/min at 10,000 G and V_max less than 2 cm/min at 500 G.

17. The solid-phase particles as in claim 15, further comprising oxides, carbides, hydroxides, nitrides, phosphates, alloys, ceramics, clays, glasses, crystals and mixtures thereof.

18. The solid-phase particles as in claim 15, wherein the biopolymer binding agent is an affinity ligand comprising organosilane, alkyl amine or PEI.

19. The solid-phase particles as in claim 18, wherein the organosilane is selected from the group consisting of trimethoxysilane, triethoxysilane or a mixture thereof.

20. The solid-phase particles as in claim 18, wherein the organosilane is an amino silane, epoxy silane, alkyl silane, or mercaptosilane.

21. The solid-phase particles as in claim 20, wherein the amino silane is selected from the group consisting of 3-aminopropyl-triethoxysilane and 3-aminopropyl-trimethoxysilane.

22. The solid-phase particles as in claim 20, wherein the epoxy silane is selected from the group consisting of 3-glycidyloxypropyltriethoxysilane and glycidyloxypropyltrimthoxysilane.

23. The solid-phase particles as in claim 19, wherein the organosilane is a styrene silane, acrylamide silane or vinylsilane.

24. The solid-phase particles as in claim 16, wherein the biopolymer binding agent is applied to the solid-phase particles up to four times forming 1st, 2^nd, 3^rd and 4^th biopolymer binding agent layers.

25. The solid-phase particles as in claim 24, wherein the 1^st layer comprises aminosilanes.

26. The solid-phase particles as in claim 24, wherein the 2^nd layer comprises polyamines.

27. The solid-phase particles as in claim 26, wherein the polyamine comprises linear or branched polymers of polyethylenimine, polylysine, proteins, polypeptides, chitins, chitosins, or a small molecule comprising diaminoethane, diaminodipropylamine, spermidine, lysine, or mixtures thereof.

28. The solid-phase particles as in claim 27, wherein the third layer is formed by reactive molecules selected from the group consisting of anhydrides, divinyl sulfone, hydrazides, triazines, carboxylates, aldehydes, haloalkyles, haloacetates, and hydroxyls.

29. The solid-phase particles as in claim 27, wherein the third layer comprises steroids or dyes.

30. The solid-phase particles as in claim 28, wherein the fourth layer is formed with amino mercaptans, polypeptides, proteins, modified nucleic acids, or oligonucleotides.

31. The solid-phase particles as in claim 28, wherein the third layer is divinyl sulfone and the fourth layer comprises thiols, mixtures of mercaptans, polypeptides, proteins, or thiophosphates.

32. The solid-phase particles as in claim 16, wherein the biopolymer is a DNA, RNA, immunoglobulin, protein or peptide.

33. The solid-phase particles as in claim 16, wherein the metals comprise transition metals.

34. The solid-phase particles as in claim 16, wherein the transition metals are lanthanides.

35. The solid-phase particles as in claim 16, wherein the semi-metals comprise Group III, Group IV, and Group V elements.

36. A method of manipulation of biomolecules comprising the step of using the solid-phase nanoparticles as in claim 16 to bind the biomolecules.

37. A method of affinity chromatography using the solid-phase particles as in claim 15, comprising interacting the solid-phase particles with a mixture of biomolecules causing adsorption of specific biomolecules.

38. A method of affinity chromatography using the solid-phase particles as in claim 36, comprising interacting the solid-phase particles with a mixture of biomolecules with the solid-phase in colloidal suspension causing adsorption of specific biomolecules in solution or suspension.

39. The method of affinity chromatography as in claim 38 wherein the separation of solid and liquid phases is achieved by centrifugation.

40. The method of affinity chromatography as in claim 39, wherein the centrifugation is achieved by using a centrifugal field strength of 10,000 G or less applied for one hour or less.

41. The method of affinity chromatography as in claim 38, wherein the separation of solid and liquid phases is achieved by filtration.

42. The method of affinity chromatography as in claim 41, wherein the filtration is achieved by using filters having pores of a dimension such that only liquid phase is filtered out retaining the solid-phase particles.

43. The method of affinity chromatography as in claim 38, wherein the separation of solid and liquid phases is achieved by flocculation.

44. The method of affinity chromatography as in claim 43, wherein the flocculation is achieved by allowing formation of particle-biomolecule complex aggregates that have a greater sedimentation rate and size that can no longer stay in suspension.

51. The method according to claim 39 is isolation of proteins or nucleic acids.

52. The method according to claim 39 is concentration of proteins or nucleic acids.

53. The method according to claim 39 is enrichment of proteins or nucleic acids.

54. The method according to claim 39, wherein the proteins or nucleic acids are in solution phase.

55. The method according to claim 39, wherein the proteins or nucleic acids are in solid-phase.

56. The method according to claim 44, wherein the proteins or nucleic acids are bound to a surface matrix.

58. The method of affinity chromatography according to claim 22, wherein the solid-phase particles in suspension are single, a mixture or a class of affinity ligands comprising peptides, avidins, immunoglobulins, lectins, or nucleic acids.

45. A method of isolating a biopolymer from a sample, the method comprising:

preparing solid-phase nanoparticles by washing,

coating the solid-phase nanoparticles with a coating,

treating the sample containing a biopolymer with the solid-phase nanoparticles,

suspending the solid-phase nanoparticles in a colloidal solution and exposing the nanoparticles to the sample solution,

separating the solid-phase nanoparticles from the sample, and

extracting the biopolymer from the solid-phase nanoparticles.

46. The method of isolating the biopolymer from a sample as in claim 45, wherein the biopolymer is selected from the group of DNA, RNA, immunoglobulin, protein and peptide.

57. The method according to claim 45, wherein the matrix is a filter membrane, fiber, solid-phase glass or plastic surface.