Method for manufacturing microarrays based on the immobilization of porous substrates on thermally modifiable surfaces

• US 20040009584A1
• Filed: 03/13/2003
• Published: 01/15/2004
• Est. Priority Date: 03/13/2002
• Status: Abandoned Application

• Current Assignee: Genomics USA, Inc.
• Original Assignee: Genomics USA, Inc.
• Inventors: Hogan, Michael E., Mitra, Rahul
• Application Number:
  US10/388,061
• Publication Number:
  US 20040009584A1
• Time in Patent Office: Days
• US Class Current: 435/287.2
• 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 2219/00459 Beads
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00646 the compounds being bound to beads immobilised on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/545 Synthetic resin

Abstract

The present invention is directed to a method of manufacturing a high density analysis device, preferably a microarray, comprising attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material; ii) immobilizing a porous or non-porous material onto the surface material; and iii) binding the immobilized porous or non-porous material with a probe to provide the high-density analysis device, and articles of same.

Claims

1. A method of manufacturing a high-density molecular analysis device comprising

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous material onto the surface material;

iii) reacting the porous material with a probe to generate an element; and

iv) repeating step iii) to provide the high-density molecular analysis device comprising a plurality of elements, wherein each element is homogeneous for the probe of that element.

2. The method of claim 1, wherein the step of reacting the porous material with the probe is performed before the step of immobilizing the porous material.

3. The method of claim 1, wherein the activatable material is characterized by having an activating temperature of less than 150°

C.

4. The method of claim 1, wherein the activatable material is a polymer.

5. The method of claim 4, wherein said polymer is agarose.

6. The method of claim 5, wherein said probe is a cell.

7. The method of claim 1, wherein the activatable material is a sol-gel glass.

8. The method of claim 1, wherein the activatable material comprises an adhesive, wherein the adhesive is thermally-cured.

9. The method of claim 8, wherein the adhesive is an epoxide, an acrylic, a wax, a resin or an organic molecule that functions as a synthetic support.

10. The method of claim 1, wherein the activatable material comprises an adhesive, wherein the adhesive is photo-cured.

11. The method of claim 10, wherein the adhesive is an epoxide or an acrylic.

12. The method of claim 1, wherein said the step of immobilizing is achieved by trapping the porous material onto the surface material, wherein the surface material is in direct contact with at least one pore of the porous material.

13. The method of claim 1, wherein said porous material is characterized by having a diameter of at least about 1 micrometer.

14. The method of claim 1, wherein said porous material comprises a synthetic polymer.

15. The method of claim 1, wherein said porous material comprises a biomolecular aggregate.

16. A method of manufacturing a high-density molecular analysis device comprising

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous or non-porous material onto the surface material;

iii) reacting the porous or non-porous material with a probe to generate an element; and

iv) repeating step iii) to provide the high-density molecular analysis device comprising a plurality of elements, wherein each element is homogeneous for the probe of that element.

17. A method of detecting a bio-molecule comprising the steps of:

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous material onto the surface material;

iii) reacting the porous material with a probe to provide an element;

iv) repeating step iii) to provide a high-density analysis device comprising a plurality of elements having a characteristic probe;

v) applying a sample comprising a bio-molecule to the high-density analysis device;

vi) binding the bio-molecule to the probe; and

v) detecting said binding.

18. The method of claim 17, wherein the step of reacting the porous material with the probe is performed before the step of immobilizing the porous material.

19. The method of claim 17, wherein the activatable material is characterized by having an activating temperature of less than 150°

C.

20. The method of claim 17, wherein the activatable material comprises an adhesive, wherein the adhesive is thermally-cured.

21. The method of claim 17, wherein said the step of immobilizing is achieved by trapping the porous material onto the surface material, wherein the surface material is in direct contact with at least one pore of the porous material.

22. The method of claim 17, wherein said porous material comprises an organism having a dimension in the range of about 1 micrometer to about 100 millimeters.

23. The method of claim 17, wherein said porous material is characterized by having a diameter of at least about 1 micrometer.

24. The method of claim 17, wherein said probe comprises a nucleic acid sequence, wherein the nucleic acid sequence is complementary to a nucleic acid sequence of the bio-molecule.

25. The method of claim 17, wherein said probe comprises a polypeptide having a complementary molecule, wherein the bio-molecule comprises the complementary molecule.

26. The method of claim 17, wherein said probe is an antibody having a complementary antigen, wherein the bio-molecule is the antigen.

27. The method of claim 17, wherein said probe is a polypeptide having a biological activity, wherein the bio-molecule alters the biological activity of the polypeptide.

28. The method of claim 17, wherein said probe is labeled with a compound that fluoresces.

29. The method of claim 28, wherein the step of detecting comprises fluorescence spectroscopy.

30. The method of claim 17, wherein said probe is a molecule that alters a biological activity of a polypeptide, wherein the bio-molecule comprises the polypeptide.

31. A method of detecting a bio-molecule comprising the steps of:

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous or non-porous material onto the surface material;

iii) reacting the porous or non-porous material with a probe to provide an element;

iv) repeating step iii) to provide a high-density analysis device comprising a plurality of elements having a characteristic probe;

v) applying a sample comprising a bio-molecule to the high-density analysis device;

vi) binding the bio-molecule to the probe; and

v) detecting said binding.

32. A method of manufacturing a high-density bio-reactor comprising the steps of:

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous material onto the surface material;

iii) reacting the porous material with a probe to generate an element;

iv) repeating step iii) to provide the high-density bio-reactor comprising a plurality of elements having a characteristic probe;

v) applying a sample comprising a target to the high-density bio-reactor, wherein an interaction between the target and the probe produces a detectable product; and

vi) detecting said product.

33. The method of claim 32, wherein the step of reacting the porous material with the probe is performed before the step of immobilizing the porous material.

34. The method of claim 32, wherein the activatable material is characterized by having an activating temperature of less than 150°

C.

35. The method of claim 32, wherein the activatable material comprises an adhesive, wherein the adhesive is thermally-cured.

36. The method of claim 32, wherein said the step of immobilizing is achieved by trapping the porous material onto the surface material, wherein the surface material is in direct contact with at least one pore of the porous material.

37. The method of claim 32, wherein said target is a polypeptide of an amino acid sequence of an enzyme.

38. The method of claim 37, wherein said probe is an organic molecule that serves as a substrate for said enzyme.

39. A method of manufacturing a high-density bio-reactor comprising the steps of:

i) attaching an activatable material to a substrate, such that the activatable material and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous or non-porous material onto the surface material;

iii) reacting the porous or non-porous material with a probe to generate an element;

iv) repeating step iii) to provide the high-density bio-reactor comprising a plurality of elements having a characteristic probe;

v) applying a sample comprising a target to the high-density bio-reactor, wherein an interaction between the target and the probe produces a detectable product; and

vi) detecting said product.

40. A method of detecting expression of a polypeptide in a sample comprising the steps of:

i) attaching an activatable material to a substrate to form a surface material, such that the activatable material and the substrate are in direct contact;

ii) immobilizing a porous material onto the surface material;

iii) reacting a unit of the porous material with a probe, wherein the unit is homogenous for the probe;

iv) repeating step iii) to provide to a high density bio-sensor comprising a plurality of units, wherein each unit is homogeneous for a probe;

v) applying the sample comprising the polypeptide to the high-density bio-sensor;

vi) binding the polypeptide to the probe of at least one unit; and

vii) detecting said binding, wherein said detection indicates expression of said polypeptide in said sample.

41. The method of claim 40, wherein the step of reacting the porous material with the probe is performed before the step of immobilizing the porous material.

42. The method of claim 40, wherein the activatable material is characterized by having an activating temperature of less than 150°

C.

43. The method of claim 40, wherein the activatable material comprises an adhesive, wherein the adhesive is thermally-cured.

44. The method of claim 40, wherein said the step of immobilizing is achieved by trapping the porous material onto the surface material, wherein the surface material is in direct contact with at least one pore of the porous material.

45. The method of claim 40, wherein said probe comprises a complementary DNA, an oligonucleotide, a chromosome, a PCR product or a gene fragment.

46. The method of claim 40, wherein said probe comprises a polypeptide.

47. The method of claim 40, wherein said probe comprises an antibody.

48. The method of claim 40, wherein said probe comprises a synthetic small molecule.

49. The method of claim 40, wherein said probe comprises a whole viral particle.

50. The method of claim 40, wherein said probe comprises a viral coat protein assembly.

51. The method of claim 40, wherein said probe comprises a cell.

52. The method of claim 40, wherein said probe comprises a whole viral particle.

53. The method of claim 40, wherein said probe comprises a viral coat protein assembly.

54. The method of claim 40, wherein said probe comprises a microorganism.

55. A method of detecting expression of a polypeptide in a sample comprising the steps of:

i) attaching an activatable material to a substrate to form a surface material, such that the activatable material and the substrate are in direct contact;

ii) immobilizing a porous or non-porous material onto the surface material;

iii) reacting a unit of the porous or non-porous material with a probe, wherein the unit is homogenous for the probe;

iv) repeating step iii) to provide to a high density bio-sensor comprising a plurality of units, wherein each unit is homogeneous for a probe;

v) applying the sample comprising the polypeptide to the high-density bio-sensor;

vi) binding the polypeptide to the probe of at least one unit; and

vii) detecting said binding, wherein said detection indicates expression of said polypeptide in said sample.

56. A method of manufacturing a high-density molecular analysis device comprising

i) attaching a molecule or cell to a substrate, such that the molecule or cell and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous material onto the surface material by thermal modification;

iii) reacting the porous material with a probe to generate an element; and

iv) repeating step iii) to provide the high-density molecular analysis device comprising a plurality of elements, wherein each element is homogeneous for the probe of that element.

57. A method of manufacturing a high-density molecular analysis device comprising

i) attaching a molecule or cell to a substrate, such that the molecule or cell and the substrate are in direct contact, to form a surface material;

ii) immobilizing a porous or non-porous material onto the surface material by thermal modification;

iii) reacting the porous or non-porous material with a probe to generate an element; and

iv) repeating step iii) to provide the high-density molecular analysis device comprising a plurality of elements, wherein each element is homogeneous for the probe of that element.