Molecular combing process for detecting macromolecules

• US 6,548,255 B2
• Filed: 06/13/2001
• Issued: 04/15/2003
• Est. Priority Date: 02/11/1994
• Status: Expired due to Term

• Current Assignee: INSTITUT PASTEUR AND CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
• Original Assignee: INSTITUT PASTEUR AND CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
• Inventors: Bensimon, Aaron, Bensimon, David, Heslot, François
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US09/879,818
• Publication Number:
  US 20020031774A1
• Time in Patent Office: 671 Days
• Field of Search: 435/6, 435/7.14, 435/91.2, 536/22.1, 536/23.1, 536/24.3, 530/350, 530/387.1
• US Class Current: 435/6.16
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/682 Signal amplification
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• 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 33/6803 : General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: Y10S 435/969 : Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 : Test strip or test slide
• 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
• CPC Class Codes: Y10S 530/811 : Peptides or proteins is immobilized on, or in, an inorganic carrier
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/145555 : Hetero-N

Abstract

The subject of the present invention is a process for aligning a macromolecule (macromolecules) on the surface S of a support, characterized in that the triple line S/A/B (meniscus) resulting from the contact between a solvent A and the surface S and a medium B is caused to move on the said surface S, the said macromolecules having a part, especially an end, anchored on the surface S, the other part, especially the other end, being in solution in the solvent A.

The subject of the present invention is also a process for detecting, measuring the intramolecular distance of, separating and/or assaying a macromolecule in a sample in which a process of alignment according to the invention is used.

Claims

1. A process for detecting a macromolecule in a sample, wherein the process comprises:

(a) providing a support having a surface, wherein the surface comprises a molecule with biological activity attached thereto and wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(b) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity;

(c) passing a meniscus over the coupled product from step (b) to align the coupled product on the surface; and

(d) detecting, either directly or indirectly, the macromolecule in the coupled product.

2. The process according to claim 1, wherein the protein comprises an antibody, an antigen, a ligand, or a ligand receptor.

3. The process according to claim 1, wherein the surface comprises an organic polymer, an inorganic polymer, a metal, a metal oxide, a sulfide, or a semiconductor element.

4. The process according to claim 1, wherein the surface comprises glass, surface-oxidized silicon, gold, graphite, molybdenum sulfide, or mica.

5. The process according to claim 1, wherein the support comprises a plate, a bead, a fiber, or a particle.

6. The process according to claim 1, wherein the surface of the support comprises an exposed reactive group having an affinity for the molecule with biological activity.

7. The process according to claim 1, wherein the surface comprises a vinyl, an amine, a carboxyl, an aldehyde, or a hydroxyl group.

8. The process according to claim 6, wherein the surface of the support comprises a substantially monomolecular layer of an organic compound and wherein the substantially monomolecular layer of the organic compound comprises:

(a) an attachment group having an affinity for the support; and

(b) an exposed group having no or little affinity for the support and the attachment group under attachment conditions, but having an affinity for the molecule with biological activity.

9. A process according to claim 6, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.

10. The process according to claim 1, wherein the protein comprises avidin or streptavidin.

11. The process according to claim 1, wherein the meniscus is passed over the coupled product by removing the surface from the sample or removing the sample from the surface.

12. The process according to claim 1, wherein the meniscus is passed over the coupled product by evaporation of the sample.

13. The process according to claim 1, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.

53. The process according to one of claims 1, 14, 25, 38, or 51, wherein the molecule with biological activity is a nucleic acid.

54. The process according to one of claims 1, 14, 25, 38, or 51, wherein the molecule with biological activity is a protein.

14. A process for detecting a macromolecule in a sample, wherein the process comprises:

(a) providing a support having a surface, wherein the surface comprises a molecule with biological activity, wherein the molecule with biological activity has been aligned on the surface by passage of a meniscus over the molecule with biological activity, and wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(b) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity; and

(c) detecting, either directly or indirectly, the macromolecule in the coupled product from step (b).

15. The process according to claim 14, wherein the protein comprises an antibody, an antigen, a ligand, or a ligand receptor.

16. The process according to claim 14, wherein the surface comprises an organic polymer, an inorganic polymer, a metal, a metal oxide, a sulfide, or a semiconductor element.

17. The process according to claim 14, wherein the surface comprises glass, surface-oxidized silicon, gold, graphite, molybdenum sulfide, or mica.

18. The process according to claim 14, wherein the support comprises a plate, a bead, a fiber, or a particle.

19. The process according to claim 14, wherein the surface of the support comprises an exposed reactive group having an affinity for the molecule with biological activity.

20. The process according to claim 14, wherein the surface comprises a vinyl, an amine, a carboxyl, an aldehyde, or a hydroxyl group.

21. The process according to claim 19, wherein the surface of the support comprises a substantially monomolecular layer of an organic compound and wherein the substantially monomolecular layer of the organic compound comprises:

(a) an attachment group having an affinity for the support; and

(b) an exposed group having no or little affinity for the support and the attachment group under attachment conditions, but having an affinity for the molecule with biological activity.

22. A process according to claim 19, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.

23. The process according to claim 14, wherein the protein comprises avidin or streptavidin.

24. The process according to claim 14, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.

25. A process for detecting a macromolecule in a sample, wherein the process comprises:

(a) providing a support having a surface;

(b) anchoring a molecule with biological activity to the surface, wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(c) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity;

(d) passing a meniscus over the coupled product from step (c) to align the coupled product on the surface; and

(e) detecting, either directly or indirectly, the macromolecule in the coupled complex.

26. The process according to claim 25, wherein the protein comprises an antibody, an antigen, a ligand receptor.

27. The process according to claim 25, wherein the surface comprises an organic polymer, an inorganic polymer, a metal, a metal oxide, a sulfide, or a semiconductor element.

28. The process according to claim 25, wherein the surface comprises glass, surface-oxidized silicon, gold, graphite, molybdenum sulfide, or mica.

29. The process according to claim 25, wherein the support comprises a plate, a bead, a fiber, or a particle.

30. The process according to claim 25, wherein the surface of the support comprises an exposed reactive group having an affinity for the molecule with biological activity.

31. The process according to claim 25, wherein the surface comprises a vinyl, an amine, a carboxyl, an aldehyde, or a hydroxyl group.

32. The process according to claim 30, wherein the surface of the support comprises a substantially monomolecular layer of an organic compound and wherein the substantially monomolecular layer of the organic compound comprises:

(a) an attachment group having an affinity for the support; and

(b) an exposed group having no or little affinity for the support and the attachment group under attachment conditions, but having an affinity for the molecule with biological activity.

33. A process according to claim 30, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.

34. The process according to claim 25, wherein the protein comprises avidin or streptavidin.

35. The process according to claim 25, wherein the meniscus is passed over the coupled product by removing the surface from the sample or removing the sample from the surface.

36. The process according to claim 25, wherein the meniscus is passed over the coupled product by evaporation of the sample.

37. The process according to claim 25, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.

38. A process for detecting a macromolecule in a sample, wherein the process comprises:

(a) providing a support having a surface;

(b) anchoring a molecule with biological activity to the surface, wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(c) passing a meniscus over the molecule with biological activity to align the molecule with biological activity on the surface;

(d) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity; and

(e) detecting, either directly or indirectly, the macromolecule in the coupled product from step (d).

39. The process according to claim 38, wherein the protein comprises an antibody, an antigen, a ligand, or a ligand receptor.

40. The process according to claim 38, wherein the surface comprises an organic polymer, an inorganic polymer, a metal, a metal oxide, a sulfide, or a semiconductor element.

41. The process according to claim 38, wherein the surface comprises glass, surface-oxidized silicon, gold, graphite, molybdenum sulfide, or mica.

42. The process according to claim 38, wherein the support comprises a plate, a bead, a fiber, or a particle.

43. The process according to claim 38, wherein the surface of the support comprises an exposed reactive group having an affinity for the molecule with biological activity.

44. The process according to claim 38, wherein the surface comprises a vinyl, an amine, a carboxyl, an aldehyde, or a hydroxyl group.

45. The process according to claim 43, wherein the surface of the support comprises a substantially monomolecular layer of an organic compound and wherein the substantially monomolecular layer of the organic compound comprises:

(a) an attachment group having an affinity for the support; and

(b) an exposed group having no or little affinity for the support and the attachment group under attachment conditions, but having an affinity for the molecule with biological activity.

46. A process according to claim 43, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.

47. The process according to claim 38, wherein the protein comprises avidin or streptavidin.

48. The process according to claim 38, wherein the meniscus is passed over the molecule with biological activity by removing the surface from the sample or removing the sample from the surface.

49. The process according to claim 38, wherein the meniscus is passed over the molecule with biological activity by evaporation of the sample.

50. The process according to claim 38, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.

51. A process for quantifying the amount of a macromolecule in a sample, comprising:

a) providing a support having a surface, wherein the surface comprises a molecule with biological activity attached thereto and wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(b) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity;

(c) passing a meniscus over the coupled product from step (b) to align the coupled product on the surface; and

(d) detecting, either directly or indirectly, the macromolecule in the coupled complex; and

(e) quantifying the amount of the macromolecule in the sample.

52. A process for quantifying the amount of a macromolecule in a sample, comprising:

(a) providing a support having a surface, wherein the surface comprises a molecule with biological activity, wherein the molecule with biological activity has been aligned on the surface by passage of a meniscus over the molecule with biological activity and wherein the molecule with biological activity comprises biotin, a protein, a nucleic acid, a lipid, or a polysaccharide;

(b) contacting the sample with the surface to couple the macromolecule in the sample to the molecule with biological activity; and

(c) detecting, either directly or indirectly, the macromolecule in the coupled product from step (b); and

(d) quantifying the amount of the macromolecule in the sample.