Highly specific surface for biological reactions having an exposed ethylenic double bond, process of using the surface, and method for assaying for a molecule using the surface

• US 5,846,724 A
• Filed: 01/28/1997
• Issued: 12/08/1998
• Est. Priority Date: 02/11/1994
• Status: Expired due to Term

• Current Assignee: Centre National De La Recherche Scientifique, Institut Pasteur
• Original Assignee: Centre National De La Recherche Scientifique, Institut Pasteur
• Inventors: Bensimon, Aaron, Bensimon, David, Heslot, Fran.cedilla.ois
• Primary Examiner(s): Horlick, Kenneth R.
• Application Number:
  US08/788,483
• Time in Patent Office: 679 Days
• Field of Search: 435/6, 435/7.1, 436/94, 436/501, 436/63, 422/104
• US Class Current: 435/6.19
• 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 present invention relates especially to a highly specific surface for biological reactions, characterized in that it contains a support having at the surface at least one essentially compact layer of an organic compound having, outside the layer, an exposed group containing an ethylenic double bond having affinity for one type of molecule with biological activity under certain reaction conditions, the other elements of the layer being essentially inaccessible for the said molecules under the said reaction conditions.

Claims

1. A highly specific surface for biological reactions comprising a support having at a surface at least one compact layer of an organic compound having, outside the layer, an a nucleic acid in solution at a pH of less than 8, wherein the other elements of the layer are essentially inaccessible to the nucleic acid, and further wherein the surface has one end of a nucleic acid sequence directly attached thereto.

2. The highly specific surface according to claim 1, wherein the support is a solid support.

3. The highly specific surface according to claim 1 or claim 2, wherein the exposed group is a vinyl group.

4. The highly specific surface according to claim 1, wherein the exposed group is an ethylenic double bond formed by deposition of a monomer or a polymer on the support, or by partial depolymerization of the surface of a polymer on the support.

5. The highly specific surface according to claim 4, wherein the polymer is a polyolefin or a polyene derivative.

6. The highly specific surface according to claim 1, wherein the compact layer of the organic compound comprises:

(A) an attachment group having affinity for the support, and

(B) the exposed group containing the ethylenic double bond, having little or no affinity for the support and the attachment group, but having the affinity for one end of a nucleic acid in solution at a pH of less than 8.

7. The highly specific surface according to claim 6, wherein the attachment between the attachment group and the support is noncovalent.

8. The highly specific surface according to claim 6, wherein the attachment between the attachment group and the support is hydrophilic or hydrophobic.

9. The highly specific surface according to claim 6, wherein the attachment group is selected from the group consisting of alkyl and halogenated alkyl.

10. The highly specific surface according to claim 6, wherein the attachment group is selected from the group consisting of --CH₃, --CF₃, --CH₂ F, and --CHF₂.

11. The highly specific surface according to claim 6, wherein the attachment between the attachment group and the support is covalent.

12. The highly specific surface according to claim 1, wherein the organic compound is cross-linked.

13. The highly specific surface according to claim 6, wherein the organic compound has an attachment group at one end and an exposed group at the other end.

14. The highly specific surface according to claim 6, wherein the attachment group is a metal alkoxide.

15. The highly specific surface according to claim 6, wherein the exposed group is linked directly to the attachment group.

16. The highly specific surface according to claim 1, wherein the support at the surface comprises a polymer, a metal, a metal oxide, a semiconductor element, an oxide of a semiconductor element, or a combination thereof.

17. The highly specific surface according to claim 1, wherein the support at the surface comprises glass or silica.

18. The highly specific surface according to claim 1, wherein the support is a plate, beads, fiber, or particles.

19. The highly specific surface according to claim 18, wherein the particles are magnetic, fluorescent, or colored.

37. A kit, comprising at least one highly specific surface according to claim 1 and a package for said highly specific surface.

38. A kit according to claim 37, also comprising a fluorescent reagent or a reagent containing beads.

39. The highly specific surface according to claim 14, wherein the metal alkoxide is silane, chlorosilane, methoxysilane, ethoxysilane, silanol, or silazane.

40. The highly specific surface according to claim 39, wherein the metal alkoxide is mono-, di-, or trichlorosilane;

mono-, di-, or triethoxysilane;

or mono, di-, or trimethoxysilane.

41. The highly specific surface according to claim 6, wherein the attachment group is of the German type.

42. The highly specific surface according to claim 41, wherein the attachment group of the German type is chlorogerman, methoxygerman, or ethoxygerman.

43. The highly specific surface according to claim 6, wherein the attachment group is a phosphate, hydroxyl, hydrazide, hydrazine, amine, amide, diazonium, pyridine, sulfate, sulfonic, carboxylic, boronic, halogen, acid halide, or aldehyde group.

44. The highly specific surface according to claim 6, wherein the exposed group is linked to the attachment group by a chain containing at least one carbon atom.

45. The highly specific surface according to claim 6, wherein the exposed group is linked to the attachment group by a chain containing 3 to 30 carbon atoms.

46. The highly specific surface according to claim 16, wherein the semiconductor oxide is a silicon oxide.

20. A process for preparing a highly specific surface for biological reactions comprising:

(A) providing a substantially monomolecular and essentially compact layer of an organic compound having at least;

(1) an attachment group having an affinity for the support, and

(2) an exposed group containing an ethylenic double bond having no or little affinity for the support and the attachment group; and

(B) directly attaching one end of a nucleic acid to the exposed group at a pH of less than 8.

21. The process according to claim 20, wherein the attached nucleic acid contains a sequence complementary to a nucleic acid sequence to be isolated from a sample.

22. The process according to claim 20, wherein the attached nucleic acid is capable of specifically recognizing and attaching a protein to be isolated from a sample.

23. The process according to either claim 21 or claim 22, wherein the attached nucleic acid or the nucleic acid to be isolated from the sample is labeled with biotin, avidin, streptavidin, or derivatives thereof.

24. A process for assaying a biological molecule in a sample, comprising:

(A) providing a highly specific surface having at least one essentially compact layer of an organic compound having, outside the layer, an exposed group containing an ethylenic double bond having affinity for one end of a nucleic acid at a pH of less than 8, the other elements of the layer being inaccessible to the nucleic acid;

(B) contacting a sample potentially containing the biological molecule with the highly specific surface containing the nucleic acid directly attached thereto; and

(C) binding the biological molecule to the nucleic acid.

25. A process according to claim 24, wherein the biological molecule is a protein, an antibody, or a nucleic acid.

26. A process according to claim 24, wherein the bound biological molecule is detected by a fluorescent or a non-fluorescent reagent.

27. A process according to claim 26, wherein the reagent is fluorescent and the surface has a fluorescence lower than fluorescence of the fluorescent reagent.

28. A process according to claim 26, wherein the reagent comprises beads.

29. A process according to any one of claims 26, 27, or 28, wherein the detection is performed by microscopy.

30. A process according to claim 24, wherein the attached nucleic acid and the sample biological molecule are differentially labeled.

32. A process according to claim 26 or claim 31, wherein the method of detecting is ELISA or RIA.

33. A process according to either claims 24 or 31, wherein the biological molecule in the sample is the product of a nucleic acid amplification.

48. A process according to any one of claims 24 or 31, wherein the biological molecule in the sample is DNA, RNA, protein, antibody, or antigen.

49. A process according to claim 48, wherein the biological molecule is DNA or RNA.

31. A process for the mapping of genes comprising:

(A) providing a highly specific surface having at least one compact layer of an organic compound having, outside the layer, an exposed group containing an ethylenic double bond directly attached to one end of a nucleic acid, the other elements of the layer being inaccessible to the nucleic acid;

(B) contacting the attached nucleic acid with a sample containing a biological molecule, where the biological molecule is a second nucleotide sequence complementary to the attached nucleic acid;

(C) hybridizing the attached nucleic acid with the second nucleotide sequence; and

(D) detecting the presence of hybridization between the attached nucleic acid and the second nucleotide sequence.

47. A process according to claim 31, wherein the position or size of the second nucleotide sequence hybridized with the attached nucleic acid is determined.

34. A process for directly attaching an end of a nucleic acid to a surface comprising:

(A) providing a highly specific surface having at least one compact layer of an organic compound having, outside the layer, an exposed group containing an ethylenic double bond having affinity for one end of a nucleic acid, the other elements of the layer being inaccessible to the nucleic acid; and

(B) directly absorbing the nucleic acid to the double bonds by contacting one end of the nucleic acid with the surface at a determined pH region.

35. A process for directly attaching a nucleic acid on a surface comprising:

(A) providing a highly specific surface having at least one compact layer of an organic compound having, outside the layer, an exposed group containing an ethylenic double bond having affinity for one end of a nucleic acid, the other elements of the layer being inaccessible to the nucleic acid; and

(B) directly absorbing the nucleic acid to the double bonds by contacting one end of the nucleic acid with the surface at a pH of less than 8.

36. A process for directly attaching an end of a nucleic acid according to claim 35, wherein the pH is between 5 and 6.

50. A process for preparing a highly specific surface for biological reactions comprising:

(A) providing a substantially monomolecular and essentially compact layer of an organic compound having at least;

(1) attachment groups having an affinity for a support, and

(2) exposed groups containing ethylenic double bonds having no or little affinity for the support and the attachment groups; and

(B) attaching a linear strand of a nucleic acid to the exposed groups at a pH of less than 8.

51. The process according to claim 50, wherein the attached nucleic acid contains a sequence complementary to a nucleic acid sequence to be isolated from a sample.

52. The process according to claim 50, wherein the attached nucleic acid is capable of specifically recognizing and attaching a protein to be isolated from a sample.

53. The process according to claim 51 or claim 52, wherein the attached nucleic acid or the nucleic acid to be isolated from the sample is labeled with biotin, avidin, streptavidin, or derivatives thereof.

54. A process for assaying a biological molecule in a sample, comprising:

(A) providing a highly specific surface having at least one essentially compact layer of an organic compound having, outside the layer, exposed groups containing ethylenic double bonds having affinity for a linear strand of a nucleic acid at a pH of less than 8, the other elements of the layer being essentially inaccessible to the nucleic acid;

(B) contacting a sample potentially containing a biological molecule with the highly specific surface having the linear strand of nucleic acid directly attached thereto; and

(C) binding the biological molecule to the attached nucleic acid.

55. A process according to claim 54, wherein the biological molecule is a protein, an antibody, or a nucleic acid.

56. A process according to claim 54, wherein the bound biological molecule is detected by a fluorescent or a non-fluorescent reagent.

57. A process according to claim 56, wherein the reagent is fluorescent and the surface has a fluorescence lower than fluorescence of the fluorescent reagent.

58. A process according to claim 56, wherein the reagent comprises beads.

59. A process according to any one of claims 56, 57, or 58, wherein the detection is performed by microscopy.

61. A process according to claim 56 or claim 60, wherein the method of detecting is ELISA or RIA.

63. A process according to claim 54 or claim 60, wherein the biological molecule in the sample is DNA, RNA, protein, antibody, or antigen.

64. A process according to claim 54 or claim 60, wherein the biological molecule in the sample is the product of a nucleic acid amplification.

60. A process for mapping a gene comprising:

(A) providing a highly specific surface having at least one essentially compact layer of an organic compound having, outside the layer, exposed groups containing ethylenic double bonds attached to a linear strand of a nucleic acid, the other elements of the layer being essentially inaccessible to the nucleic acid;

(B) contacting the attached nucleic acid with a sample containing a biological molecule, where the biological molecule is a second nucleotide sequence complementary to the attached nucleic acid;

(C) hybridizing the attached nucleic acid with the second nucleotide sequence; and

(D) detecting the presence of hybridization between the attached nucleic acid and the second nucleotide sequence;

wherein the nucleic acid is directly attached to the surface.

62. A process according to claim 60, wherein the position or size of the second nucleotide sequence hybridized with the attached nucleic acid is determined.

65. A process for attaching a nucleic acid on a surface comprising:

(A) providing a highly specific surface having at least one essentially compact layer of an organic compound having, outside the layer, exposed groups containing ethylenic double bonds having affinity for a linear strand of a nucleic acid at a pH of less than 8, the other elements of the layer being essentially inaccessible to the nucleic acid; and

(B) absorbing the nucleic acid directly to the surface by contacting the nucleic acid with the surface at a pH of less than 8.

66. A process according to claim 65, wherein the pH is between 5 and 6.