Interaction detection on several probe arrays
First Claim
1. A probe array for qualitative and/or quantitative detection of target molecules in a sample by molecular interactions between probe molecules and target molecules on the probe array, comprising an array surface and probe molecules immobilised on the array surface at defined sites, wherein the probe molecules have at least one label and at least one selectively cleavable bond between the site of their immobilisation on the array surface and the label.
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Accused Products
Abstract
The invention concerns a method for detecting interactions between probe molecules and target molecules, whereby marking target molecules is not required. The invention further concerns probe arrays and kits suited to such a method, as well as a method for production, quality control and standardization of probe arrays.
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Citations
61 Claims
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1. A probe array for qualitative and/or quantitative detection of target molecules in a sample by molecular interactions between probe molecules and target molecules on the probe array, comprising an array surface and probe molecules immobilised on the array surface at defined sites,
wherein the probe molecules have at least one label and at least one selectively cleavable bond between the site of their immobilisation on the array surface and the label.
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26. A method for producing a probe array, comprising:
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a) synthesizing probe molecules having a label and having a selectively cleavable bond between the site of their immobilisation on the array surface and the label; and
b) immobilizing the probe molecules via a defined position within the probe molecules at specific sites on the array surface.
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27. A method for producing an array of probes on an array surface by in situ synthesis of the probe molecules on predetermined positions of the array surface, comprising:
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a) providing an array surface which can be activated by suitable reagents or is provided with protective groups;
b) coupling or immobilising subunits of the probe molecules to be synthesised to predetermined sites on the array surface; and
c) synthesizing the probe molecules in situ following said coupling or immobilizing by incorporation of a label and a selectively cleavable bond between the site of the immobilisation of the probe molecules on the array surface and the label. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35)
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59. A method for the production of monomer building blocks suitable for DNA synthesis, which can be used for the formation of a labile bond in probe molecules, comprising:
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a) esterifying the 5′
-OH group of a nucleoside with an acid suitable as leaving group, to produce an ester;
b) reacting the ester with a thioester;
c) saponifying the thioester to form a thiol;
d) protecting the thiol function with protective groups suitable for the phosphotriester or phosphoramidite method; and
e) activating the protected thiol at the 3′
position using the phosphotriester or phosphoramidite method.
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60. A method for the production of monomer building blocks suitable for DNA synthesis, which can be used for the formation of a labile bond in probe molecules, comprising:
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a) reacting a compound suitable as a protective group for the phosphotriester or phosphoramidite method to form a thiol;
b) esterifying the 5′
-OH group of a nucleoside with an acid suitable as leaving group, to form an ester;
c) reacting the thiol of a) with the ester of b); and
d) activating the protected thiol at the 3′
position using the phosphotriester or phosphoramidite method.
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61. The compound 5′
- -S-(dimethoxytrityl)-mercapto-5′
-deoxynucleoside-3′
-O-(2-cyanoethyl, N,N′
-diisopropyl-phosphite.
- -S-(dimethoxytrityl)-mercapto-5′
Specification