Nanoparticles for manipulation of biopolymers and methods of thereof

• US 7,964,380 B2
• Filed: 01/23/2006
• Issued: 06/21/2011
• Est. Priority Date: 01/21/2005
• Status: Expired due to Fees

• Current Assignee: Argela Technologies (Levent Yapilandirma Yönetimi Telekomünikasyon AS)
• Original Assignee: Argela Technologies (Levent Yapilandirma Yönetimi Telekomünikasyon AS)
• Inventors: Hogan, Michael E., Diggins, Paul, Utermohlen, Joseph G.
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US11/338,124
• Publication Number:
  US 20060177855A1
• Time in Patent Office: 1,975 Days
• Field of Search: 435/177, 435/174, 435/6
• US Class Current: 435/177
• 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, immobilization and archival storage of biopolymers is disclosed.

Claims

1. A method of manipulating biomolecules in a sample comprising the steps of:

a. providing solid-phase, non-porous particles having a surface area to volume ratio greater than 10, and a density greater than 2;

b. selecting a subset of the solid-phase, non-porous particles having sedimentation rates in water where the sedimentation velocity is between about 0.1 cm/min at 10,000 G (Vmin) and about 2 cm/min at 500 G (Vmax) at standard temperature and pressure, and wherein the particles form a colloidal suspension in aqueous solution;

c. surface-modifying the subset of the solid-phase, non-porous particles to alter their affinity for one or more classes of biomolecules, wherein the step of surface-modifying the subset of solid-phase, non-porous particles further comprises activating the subset of solid-phase, non-porous particles, passivating the subset of solid-phase, non-porous particles, or combinations thereof;

d. incubating the sample with the surface-modified particles, thereby forming a plurality of surface-modified particle-biomolecule complexes;

wherein the formation of the plurality of surface-modified particle-biomolecule complexes is reversible; and

e. separating the plurality of surface-modified particle-biomolecule complexes from the sample.

2. The method of claim 1 further comprising the steps of:

f. disassociating the plurality of surface-modified particle-biomolecule complexes thereby releasing the biomolecules from the surface-modified particles; and

g. isolating the biomolecules.

3. The method of claim 1 wherein the method of manipulating biomolecules further comprises a step of isolating the biomolecules from the sample.

4. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 2 and about 2.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 60 nm to about 1000 nm.

5. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 2.5 and about 3, and effective spherical diameters as determined by two times Stokes radius in the range of about 40 nm to about 800 m.

6. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 3 and about 3.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 35 nm to about 400 nm.

7. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 3.5 and about 4, and effective spherical diameters as determined by two times Stokes radius in the range of about 20 nm to about 700 nm.

8. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 4 and about 4.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 30 nm to about 600 nm.

9. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 4.5 and about 5, and effective spherical diameters as determined by two times Stokes radius in the range of about 25 nm to about 550 nm.

10. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 5 and about 5.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 25 nm to about 500 nm.

11. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 5.5 and about 6, and effective spherical diameters as determined by two times Stokes radius in the range of about 25 nm to about 450 nm.

12. The method of claim 1 wherein solid-phase, non-porous particles have a density between about 6 and about 6.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 20 nm to about 450 nm.

13. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 6.5 and about 7, and effective spherical diameters as determined by two times Stokes radius in the range of about 20 nm to about 400 nm.

14. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 7 and about 7.5, and effective spherical diameters as determined by two times Stokes radius in the range of about 20 nm to about 400 nm.

15. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 7.5 and about 14, and effective spherical diameters as determined by two times Stokes radius in the range of about 15 nm to about 300 nm.

16. The method of claim 1 wherein the solid-phase, non-porous particles have a density between about 14 and about 20, and effective spherical diameters as determined by two times Stokes radius in the range of about 12 nm to about 240 nm.

17. The method of claim 1 wherein the solid-phase, non-porous particles further comprise a material selected from the group consisting of oxides, carbides, hydroxides, nitrides, phosphates, alloys, ceramics, clays, glasses, crystals, and mixtures thereof.

18. The method of claim 1 wherein the solid-phase, non-porous particles further comprise of metals or semi-metals.

19. The method of claim 18 wherein the metals are selected from the group consisting of Al, Ca, Co, Cu, Fe, Ga, Mg, Mn, In, Ni, Ti, and Zn.

20. The method of claim 17 wherein the step of surface modifying the subset of solid-phase, non-porous particles further comprises passivating the subset of solid-phase, non-porous particles.

21. The method of claim 20 wherein the step of passivating the subset of solid-phase, non-porous particles further comprises passivating with at least one anion selected from the group consisting of borate, phosphate, fluoride, and combinations thereof.

22. The method of claim 1 wherein the step of activating the subset of solid-phase, non-porous particles further comprises coating the subset of solid-phase, non-porous particles with organosilane, alkyl amine, polyethylenimine (PEI), or combinations thereof.

23. The method of claim 22 wherein the organosilane further comprises an amino silane, an epoxy silane, an alkyl silane, a mercaptosilane, a styrene silane, an acrylamide silane, a vinyl silane or combinations thereof.

24. The method of claim 23 wherein the alkyl silane is selected from the group consisting of trimethoxysilane, triethoxysilane, and mixtures thereof.

25. The method of claim 23 wherein the amino silane is selected from the group consisting of 3-aminopropyl-triethoxysilane, 3-aminopropyl-trimethoxysilane, and combinations thereof.

26. The method of claim 23 wherein the epoxy silane is selected from the group consisting of 3-glycidyloxypropyltriethoxysilane, glycidyloxypropyltrimthoxysilane, and combinations thereof.

27. The method of claim 1 wherein the step of activating the subset of solid-phase, non-porous particles further comprises coating the subset of solid-phase, non-porous particles with one or more biopolymer binding agents specific for one or more classes of biomolecules.

28. The method of claim 27 wherein the step of coating the subset of solid-phase, non-porous particles with the one or more biopolymer binding agents further comprises a step of applying more than one coating of the one or more biopolymer binding agents to the subset of solid-phase, non-porous particles, thereby forming multiple layers of biopolymer binding agents on the surface of subset of solid-phase, non-porous the particles.

29. The method of claim 28 wherein the step of applying more than one coating of the one or more biopolymer binding agents further comprises a step of applying a first layer comprising an aminosilane.

30. The method of claim 28 wherein the step of applying more than one coating of the one or more biopolymer binding agents further comprises a step of applying a second layer comprising a polyamine.

31. The method of claim 30 wherein the polyamine further comprises linear or branched polymers of polyethylenimine, polylysine, proteins, polypeptides, chitins, chitosins, diaminoethane, diaminodipropylamine, spermidine, lysine, or combinations thereof.

32. The method of claim 28 wherein the one or more biopolymer binding agents further comprise reactive molecules selected from the group consisting of anhydrides, divinyl sulfones, hydrazides, triazines, carboxylates, aldehydes, haloalkyles, haloacetates, hydroxyls, and combinations thereof.

33. The method of claim 28 wherein the multiple layers of biopolymer binding agents comprise at least one layer further comprising of steroids or dyes.

34. The method of claim 28 wherein the multiple layers of biopolymer binding agents comprise at least one layer further comprising of amino mercaptans, polypeptides, proteins, modified nucleic acids, oligonucleotides, or combinations thereof.

35. The method of claim 1 wherein the step of separating the plurality of surface-modified particle-biomolecule complexes from the sample further comprises a step of separation by centrifugation.

36. The method of claim 35 wherein the centrifugation is carried out by applying a centrifugal field strength of 10,000 G or less applied for one hour or less.

37. The method of claim 1 wherein the solid-phase, non-porous particles further comprise kaolin.

38. The method of claim 37 wherein the step of surface-modifying the subset of solid-phase, non-porous particles comprises a step of surface passivating the subset of solid-phase, non-porous particles with borate, phosphate, fluoride, or combinations thereof.

39. The method of claim 37 wherein the step of surface-modifying the subset of solid-phase, non-porous particles comprises a step of coating the subset of solid-phase, non-porous particles with one or more biopolymer binding agents.

40. The method of claim 39 wherein the one or more biopolymer binding agents further comprise organosilane, alkyl amine, polyethylenimine (PEI), or combinations thereof.

41. The method of claim 1 further comprising purification of the biomolecules.

42. The method of claim 1 further comprising a step of storing the biomolecules.

43. The method of claim 2 wherein the surface-modified particles further comprise polyethylenimine-epoxysilane coated kaolin.

44. The method of claim 2 wherein the surface-modified particles further comprise phosphate treated kaolin.

45. The method of claim 2 wherein the surface-modified particles further comprise epoxy-dithiothreitol-divinylsulfone coated particles.

46. The method of claim 2 wherein the surface-modified particles further comprise amino-silane coated kaolin.

47. The method of claim 2 wherein the surface-modified particles further comprise metal oxide particles.

48. The method of claim 2 wherein the surface-modified particles further comprise amino-silane coated metal oxide particles.

49. The method of claim 47 wherein the metal oxide particles are selected from the group consisting of aluminum oxide, titanium oxide, tungsten oxide, and zirconium oxide.

50. The method of claim 47 wherein the metal oxide particles are coated with borate.

51. The method of claim 47 wherein the metal oxide particles are coated with phosphate.

52. The method of claim 47 wherein the metal oxide particles are coated with inosine monophosphate.

53. The method of claim 2 wherein the surface-modified particles further comprise borate treated kaolin.

54. The method of claim 2 wherein the step of surface-modifying the subset of solid-phase, non-porous particles further comprises coating the subset of solid-phase, non-porous particles with a thiophilic epoxy-silane-dithiothreitol-divinylsulfone ligand.

57. The method of claim 20 wherein the step of passivating the subset of solid-phase, non-porous particles comprises passivating with one or more compounds selected from the group consisting of silicate, borate, phosphate, sulfate, carbonate, arsenate, vanadate, permanganate, molybdate, tungstenate, chromate, fluoride, EDTA, EGTA, citrate, oxalate, acetate, formate, and combinations thereof.

55. A method of storing biomolecules comprising the steps of:

a. providing a sample containing biomolecules;

b. providing solid-phase, non-porous particles having a surface area to volume ratio greater than 10, and a density greater than 2;

c. selecting a subset of the solid-phase, non-porous particles having sedimentation rates in water where the sedimentation velocity is between about 0.1 cm/min at 10,000 G (Vmin) and about 2 cm/min at 500 G (Vmax) at standard temperature and pressure, and wherein the particles form a colloidal suspension in aqueous solution;

d. surface-modifying the subset of the solid-phase, non-porous particles to alter their affinity for one or more classes of biomolecules, wherein the step of surface-modifying the subset of solid-phase, non-porous particles further comprises activating the subset of solid-phase, non-porous particles, passivating the subset of solid-phase, non-porous particles, or combinations thereof;

e. incubating the sample with the surface-modified particles, thereby forming a plurality of surface-modified particle-biomolecule complexes;

wherein the formation of the plurality of surface-modified particle-biomolecule complexes is reversible;

f. isolating the plurality of surface-modified particle-biomolecule complexes from the sample; and

g. storing the plurality of surface-modified particle-biomolecule complexes.

56. The method of claim 55 further comprising the steps of:

h. disassociating the stored plurality of surface-modified particle-biomolecule complexes thereby releasing the biomolecules from the surface-modified particles; and

i. isolating the biomolecules.