METHODS AND DEVICES FOR BIOMOLECULAR ARRAYS

• US 20100120630A1
• Filed: 01/30/2008
• Published: 05/13/2010
• Est. Priority Date: 01/30/2007
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Barbee, Kristopher D., Huang, Xiaohua
• Granted Patent: US 9,063,133 B2
• Time in Patent Office: Days
• US Class Current: 506/13
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00655 the compounds being bound to magnets embedded in or on the solid supports
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G03F 7/00 Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor using photoresist structures for special production processes, see the relevant places, e.g. B44C, H01L, e.g. H01L21/00, H05K
• CPC Class Codes: G03F 7/16 Coating processes; Apparatus therefor applying coatings to base materials in general B05; applying photosensitive compositions to base for photographic purposes G03C1/74

Abstract

The disclosure provides methods and devices for rapidly assembling high density biomolecular arrays.

Claims

1. A method for rapidly assembling a high density biomolecular array, the method comprising the steps of:

a) patterning an array of wells into photoresist on a surface by photolithography;

b) assembling a suspension of particles into the array of wells;

c) immobilizing the particles to the surface via affinity binding; and

d) removing the unbound suspension of particles and photoresist from the surface to provide the high density biomolecular array.

2. The method of claim 1, wherein the suspension of particles conjugated to a biomolecule is a suspension of superparamagnetic microbeads conjugated to a biomolecule, wherein the superparamagnetic microbeads are assembled into the array of wells using a magnet.

3. The method of claim 1, wherein the surface is a glass surface.

4. The method of claim 3, wherein the glass surface is derivatized with silane and biotin, and/or avidin and/or streptavidin.

5. The method of claim 4, wherein the glass surface is spin-coated with photoresist.

6. The method of claim 5, wherein the photoresist is Microposit S1805 photoresist.

7. The method of claim 6, wherein the array of wells are patterned by photolithography using a 0.2 second exposure to 365 nM light (˜

475 mW/cm²) through a chrome-on-quartz photomask using a wafer stepper system (GCA Autostep

     200) equipped with an Olympus 2145 lens (5×

reduction/0.45 NA).

8. The method of claim 7, wherein the size of the array of wells varies with the exposure and chemical etching parameters of the photolithography technique.

9. The method of claim 8, wherein the array of wells has a density of about 100 million wells per square centimeter.

10. The method of claim 9, wherein the array of wells has a parabolic geometry wherein the top of a well is about 600 nm in diameter and the bottom of a well is about 250 nm in diameter.

11. The method of claim 2, wherein the superparamagnetic microbeads are coated with biotin, and/or avidin and/or streptavidin.

12. The method of claim 11, wherein the superparamagnetic microbeads are suspended in a buffer.

13. The method of claim 12, wherein the assembly of the superparamagnetic microbeads into the array of wells occurs in a flow cell.

14. The method of claim 13, wherein the superparamagnetic microbeads are assembled into the array of wells using a neodymium iron boron magnet.

15. The method of claim 14, wherein each well contains a single superparamagnetic microbead.

16. The method of claim 15, wherein the superparamagnetic microbeads are immobilized in the array of wells via biotin-avidin and/or biotin-streptavidin affinity binding.

17. The method of claim 2, wherein the high density biomolecular array is a high density protein array, or a high density DNA array of biomolecules.

18. A high density biomolecular array prepared by the method of claim 1.

19. A high density biomolecular array prepared by the method of claim 2.

20. The method of claim 1, wherein the suspension of particles conjugated to a biomolecule is a suspension nanoparticles conjugated to a biomolecule.

21. The method of claim 20, wherein the surface is a glass surface.

22. The method of claim 21, wherein the glass surface is derivatized with silane and biotin, and/or avidin and/or streptavidin.

23. The method of claim 22, wherein the glass surface is spin-coated with photoresist.

24. The method of claim 23, wherein the photoresist is Microposit S1805 photoresist.

25. The method of claim 24, wherein the array of wells are patterned by photolithography using a 0.2 second exposure to 365 nM light (˜

475 mW/cm²) through a chrome-on-quartz photomask using a wafer stepper system (GCA Autostep

     200) equipped with an Olympus 2145 lens (5×

reduction/0.45 NA).

26. The method of claim 25, wherein the size of the array of wells varies with the exposure and chemical etching parameters of the photolithography technique.

27. The method of claim 26, wherein the array of wells has a density of about 100 million wells per square centimeter.

28. The method of claim 27, wherein the array of wells has a parabolic geometry wherein the top of a well is about 600 nm in diameter and the bottom of a well is about 250 nm in diameter.

29. The method of claim 20, wherein the nanoparticles are coated with biotin, and/or avidin and/or streptavidin.

30. The method of claim 29, wherein the nanoparticles are suspended in a buffer.

31. The method of claim 30, wherein the assembly of the nanoparticles into the array of wells occurs in a flow cell.

32. The method of claim 31, wherein the nanoparticles self assemble into the array of wells.

33. The method of claim 32, wherein each well contains a single nanoparticle.

34. The method of claim 33, wherein the nanoparticles are immobilized in the array of wells via biotin-avidin and/or biotin-streptavidin affinity binding.

35. The method of claim 20, wherein the high density nanoarray of biomolecules is a high density nanoarray of protein, or a high density nanoarray of DNA of biomolecules.

36. A high density nanoarray of biomolecules prepared by the method of claim 20.

37. The method of claim 20, wherein the surface is derivatized with a biotin-derivatized and/or amine-derivatized cross-linked polyacrylamide polymer, wherein the cross-linked polyacrylamide polymer is coated with photoresist.

38. The method of claim 37, wherein the surface is a glass surface.

39. The method of claim 38, wherein the glass surface is coated with a biotin-derivatized cross-linked polyacrylamide polymer.

40. The method of claim 39, wherein the biotin-derivatized cross-linked polyacrylamide polymer is spin-coated with photoresist.

41. The method of claim 40, wherein the photoresist is Microposit S1805 photoresist.

42. The method of claim 41, wherein the array of wells are patterned by photolithography using a 0.2 second exposure to 365 nM light (˜

475 mW/cm²) through a chrome-on-quartz photomask using a wafer stepper system (GCA Autostep

     200) equipped with an Olympus 2145 lens (5×

reduction/0.45 NA).

43. The method of claim 42, wherein the size of the array of wells varies with the exposure and chemical etching parameters of the photolithography technique.

44. The method of claim 43, wherein the array of wells has a density of about 100 million wells per square centimeter.

45. The method of claim 44, wherein the array of wells has a parabolic geometry wherein the top of a well is about 600 nm in diameter and the bottom of a well is about 250 nm in diameter.

46. The method of claim 37, wherein the nanoparticles are coated with biotin, and/or avidin and/or streptavidin.

47. The method of claim 46, wherein the nanoparticles are suspended in a buffer.

48. The method of claim 47, wherein the assembly of the nanoparticles into the array of wells occurs in a flow cell.

49. The method of claim 48, wherein the nanoparticles are assembled into the array of wells by an electric field.

50. The method of claim 49, wherein each well contains a single nanoparticle.

51. The method of claim 49, wherein the nanoparticles are immobilized in the array of wells via biotin-avidin and/or biotin-streptavidin affinity binding.

52. The method of claim 37, wherein the high density nanoarray of biomolecules is a high density nanoarray of protein, or a high density nanoarray of DNA of biomolecules.

53. A high density nanoarray of biomolecules prepared by the method of claim 37.

54. An automated system for rapidly assembling a high density biomolecular array, comprising a microfluidic flowcell containing an array of wells on a chemically derivatized surface coated with photoresist;

a suspension of superparamagnetic microbeads conjugated to a biomolecule, and a magnet, wherein the superparamagnetic microbeads are assembled into the array of wells using the magnet.