Molecular combing process for detecting macromolecules
First Claim
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.
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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.
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Citations
54 Claims
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1. A process for detecting a macromolecule in a sample, wherein the process comprises:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 53, 54)
(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.
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9. A process according to claim 6, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.
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10. The process according to claim 1, wherein the protein comprises avidin or streptavidin.
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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.
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12. The process according to claim 1, wherein the meniscus is passed over the coupled product by evaporation of the sample.
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13. The process according to claim 1, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.
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53. The process according to one of claims 1, 14, 25, 38, or 51, wherein the molecule with biological activity is a nucleic acid.
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54. The process according to one of claims 1, 14, 25, 38, or 51, wherein the molecule with biological activity is a protein.
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14. A process for detecting a macromolecule in a sample, wherein the process comprises:
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(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). - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
(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.
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22. A process according to claim 19, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.
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23. The process according to claim 14, wherein the protein comprises avidin or streptavidin.
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24. The process according to claim 14, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.
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25. A process for detecting a macromolecule in a sample, wherein the process comprises:
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(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. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37)
(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.
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33. A process according to claim 30, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.
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34. The process according to claim 25, wherein the protein comprises avidin or streptavidin.
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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.
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36. The process according to claim 25, wherein the meniscus is passed over the coupled product by evaporation of the sample.
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37. The process according to claim 25, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.
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38. A process for detecting a macromolecule in a sample, wherein the process comprises:
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(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). - View Dependent Claims (39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50)
(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.
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46. A process according to claim 43, wherein the exposed reactive group is an ethylenic double bond, a vinyl group, or an amine group.
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47. The process according to claim 38, wherein the protein comprises avidin or streptavidin.
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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.
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49. The process according to claim 38, wherein the meniscus is passed over the molecule with biological activity by evaporation of the sample.
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50. The process according to claim 38, further comprising subjecting the coupled product to a stress to destroy any mismatches before detecting the macromolecule.
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51. A process for quantifying the amount of a macromolecule in a sample, comprising:
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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.
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52. A process for quantifying the amount of a macromolecule in a sample, comprising:
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(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.
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Specification