DEPOSITION OF METAL OXIDES ONTO SURFACES AS AN IMMOBILIZATION VEHICLE FOR CARBOXYLATED OR PHOPHATED PARTICLES OR POLYMERS

• US 20100086927A1
• Filed: 07/23/2009
• Published: 04/08/2010
• Est. Priority Date: 07/23/2008
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Fry, George A., Menchen, Steven M., Taing, Meng, Inman, Christina E.
• Granted Patent: US 8,173,198 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C07F 9/093 Polyol derivatives esterified at least twice by phosphoric acid groups
• CPC Class Codes: C07F 9/6596 having atoms other than oxygen, sulfur, selenium, tellurium, nitrogen or phosphorus as ring hetero atoms
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: Y10S 530/81 Carrier - bound or immobilized peptides or proteins and the preparation thereof, e.g. biological cell or cell fragment as carrier

Abstract

Intermediates and methods for forming activated metal complexes bound to surfaces on oxide layers, immobilizing beads to the modified surface and articles produced thereby are described. Hydroxyl groups on the oxide surfaces are reacted with a metal reagent complex of the formula Y(L-Pol)_m, where Y is a transition metal, magnesium or aluminum, L is oxygen, sulfur, selenium or an amine, and “Pol” represents a passivating agent such as a methoxyethanol, a polyethylene glycol, a hydrocarbon, or a fluorocarbon. The resulting modified surface can be further reacted with a passivating agent having a phosphate functional group or a plurality of functional groups that are reactive with or that form complexes with Y. The metal oxide surfaces exhibit minimal binding to bio-molecules, exhibit uniform deposition and immobilization of beads at high density, can be subsequently modified to create surfaces having a variety of properties, and can be used for nucleic acid sequencing and other analyses and in single-molecule detection schemes.

Claims

1. A method of immobilizing a particle on to a surface comprising:

depositing a metal oxide layer onto the surface; and

contacting a functionalized particle onto the metal oxide surface;

wherein the particle is immobilized on the surface.

2. The method of claim 1, wherein the functionalized particle comprises phosphated moieties, carboxylated moieties or a combination thereof.

3. The method of claim 2, wherein the phosphated moieties are derived from phosphodiester linkages selected from the group consisting of a nucleic acid, an oligonucleotide or a biomolecule containing nucleic acids, carboxylates, phosphonates or phosphates.

4. The method of claim 1, wherein the particle is selected from the group consisting of a bead, a refractory bead or a ligation bead.

5. The method of claim 1, wherein the surface is a glass, a metal, a metal oxide or an organic polymer.

6. The method of claim 1, wherein the particle is composed of a material selected from the group consisting of organic polymers, silicate polymers, alumina, titania, zirconia, and combinations thereof.

7. The method of claim 5, wherein the polymer is solid at operating temperatures.

8. The method of claim 5, wherein the polymer is optically transparent.

9. The method of claim 5, wherein the polymer is stable in a variety of solvents.

10. The method of claim 5, wherein the polymer is activated to comprise transition metal reactive groups.

11. The method of claim 10, wherein the reactive group is selected from hydroxyl, carboxyl, phosphate, phosphonate, silanol, alumina and oxides of transition metals.

12. The method of claim 1, wherein the surface comprises a material selected from the group consisting of silicon, aluminum, germanium, gallium, titanium, tantalum, indium, zirconium, magnesium, tin, gold, silver, hafnium, carbon and an organic polymer derivative.

13. The method of claim 5, wherein the polymer is selected from the group consisting of cyclic olefin polymer (COP), cyclic olefin co-polymer (COC), polypropylene, polycarbonate, fluorinated polyethylene, polymethyl methacrylate, polystyrene, polybutadiene, and blends thereof.

14. The method of claim 13, wherein the blend is a mixture of polystyrene, polypropylene and polybutadiene.

15. The method of claim 1 further comprising passivating the surface.

16. The method of claim 15, wherein the passivating comprises a passivating agent selected from the group consisting of poly(vinyl phosphoric acid), pyrophosphate, poly(acrylic acid), polyvinyl alcohol), polyethylene glycol comprising functionalities of poly-phosphate-, poly-carboxylate-, or poly-hydroxyl- or combinations thereof and poly(N,N-dimethylacrylamide).

25. The method of claim 1, wherein the metal oxide comprises an organic polymer derivative.

32. The method of claim 1, wherein the particle is at least 0.5 to 10 microns in size.

17. A method of making a particle immobilized on a surface comprising:

a. layering an organic soluble metal complex on the surface, forming a functionalized metal complex surface;

b. hydrolyzing the functionalized metal complex;

c. contacting a particle to the metal complex layer, wherein the bead is immobilized to the surface.

18. The method of claim 17, further comprising passivating the surface.

19. The method of claim 17, further comprising pre-treating the surface prior to layering the surface with the metal complex, wherein pre-treating comprises treating the surface with an oxygen plasma or contacting the surface with tetrabutyl ammonium hydroxide, chromic acid in sulfuric acid, potassium hydroxide in methanol, hydrogen peroxide in sulfuric acid, nitric acid in sulfuric acid, hydrogen peroxide in ammonia, sulfuric acid, hydrofluoric acid, EDTA, or combinations thereof.

20. The method of claim 18, wherein the passivating comprises a passivating agent selected from the group consisting of poly(vinyl phosphoric acid), pyrophosphate, poly(acrylic acid), poly(vinyl alcohol), polyethylene glycol comprising functionalities of poly-phosphate-, poly-carboxylate-, or poly-hydroxyl- or combinations thereof, and poly(N,N-dimethylacrylamide).

21. The method of claim 17, wherein the surface comprises a metal selected from the group consisting of silicon, aluminum, germanium, gallium, titanium, tantalum, indium, zirconium, magnesium, tin, gold, silver, hafnium, carbon, and an organic polymer derivative.

26. The method of claim 17, wherein the metal oxide comprises an organic polymer derivative.

22. A compound of the formula:

Y(L-Pol)_m

wherein Y is silicon, aluminum, germanium, gallium, titanium, tantalum, indium, hafnium, zirconium, magnesium, tin, gold, silver or another transition metal;

L is oxygen, sulfur, selenium or an amine;

each “

Pol”

group independently represents a methoxyethariol, polyethylene glycol, a substituted polyethylene glycol, a hydrocarbon, as substituted hydrocarbon, a fluorocarbon or a substituted fluorocarbon, and

m is an integer.

23. The compound of claim 22, wherein Y is selected from the group consisting of Ta, Ti, Zr, and Al.

24. The method of claim 22, further comprising:

contacting the modified surface with a plurality of functional groups that are reactive with Y or that form complexes with Y and are selected from the group consisting of;

hydroxyl groups, amine groups, phosphate groups, phosphonate groups, thiol groups, alkylphosphate groups, carboxyl groups and combinations thereof.

27. A solid support having a surface treated with a multivalent passivating agent comprising:

a. a metal oxide layer;

b. at least one functionalized particle in contact with the metal oxide layer; and

c. the passivating agent in contact with the metal oxide layer,

wherein the surface is passivated with a passivating agent.

28. The solid support of claim 27, wherein the surface comprises a substrate comprising a glass, a metal, a metal oxide or an organic polymer.

29. The solid support of claim 27, wherein the functionalized particle comprises phosphated moieties, carboxylated moieties or a combination thereof.

30. The solid support of claim 28, wherein the phosphated moieties are derived from phosphodiester linkages selected from the group consisting of a nucleic acid, an oligonucleotide or a biomolecule containing nucleic acids, carboxylates, phosphonates or phosphates.

31. The solid support of claim 27, wherein the passivating comprises a passivating agent selected from the group consisting of poly(vinyl phosphoric acid), pyrophosphate, poly(acrylic acid), poly(vinyl alcohol), polyethylene glycol comprising functionalities of poly-phosphate-, poly-carboxylate-, or poly-hydroxyl- or combinations thereof, and poly(N,N-dimethylacrylamide).

33. A surface of a substrate comprising a nucleic acid coated immobilized bead formed by the process of:

a. applying a metal oxide layer;

b. binding at least one functionalized nucleic acid coated bead with the metal oxide layer;

wherein the at least one nucleic acid coated bead is immobilized to the surface.

34. The surface comprising the immobilized bead of claim 33, wherein the surface comprises a substrate comprising a glass, a metal, a metal oxide or an organic polymer.

35. The surface comprising the immobilized bead of claim 33, wherein the functionalized nucleic acid coated bead comprises phosphated moieties, carboxylated moieties or a combination thereof.

36. The surface comprising the immobilized bead of claim 35, wherein the phosphated moieties are derived from phosphodiester linkages selected from the group consisting of a nucleic acid, an oligonucleotide or a biomolecule containing nucleic acids, carboxylates, phosphonates or phosphates.

37. The surface comprising the immobilized bead of claim 33 optionally comprising applying a passivating agent subsequent to the binding of the at least one functionalized nucleic acid coated bead.

38. The surface comprising the immobilized bead of claim 37, wherein the passivating comprises a passivating agent selected from the group consisting of poly(vinyl phosphoric acid), pyrophosphate, poly(acrylic acid), poly(vinyl alcohol), polyethylene glycol comprising functionalities of poly-phosphate-, poly-carboxylate-, or poly-hydroxyl- or combinations thereof, and poly(N,N-dimethylacrylamide).