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EP0152886A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Immobilized nucleic acid-containing probes

Assignee: Molecular Diagnostics Inc
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Filing: 1985-02-11
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Granted: —
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Expires: 2005-02-11
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Abstract

Abstract An immobilized nucleic acid-containing probe suitable for accurate hybridization gene analyses is produced by enzymatically coupling a nucleotide to a predetermined end of the nucleic acid and then selectively coupling a suitably prepared solid support, such as cellulose, through the nucleotide. Thus the ribose of the nucleotide can be oxidized by sodium periodate to form aldehyde groups which will react with a primary amino group introduced into collulose, forming a Schiff's base. For grater stability of the immobilizing bond the Schiff's base is reduced to a saturated secondary amino group by reduction with borohydride. Alternatively, the nucleotide may carry an SH or Hg moiety which will selectively react with complementary moieties on the solid support.

Inventors

  • 👤 Nanibhushan Dattagupta

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-10-29 10:13
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/544; G01N33/548; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6834

Claims Analysis

Total Claims: 18
Claim 1
Independent Claim 23 words
An immobilized nucleic acid containing probe characterized in that the nucleic acid is attached to a solid support by a single chemical linkage.
Claim 2
Dependent Claim 23 words
An immobilized nucleic acid containing probe characterized in that the nucleic acid is attached to a solid support by a single chemical linkage.
Claim 3
Dependent Claim 20 words
A probe according to claim 1, characterized in that the chemical linkage to the solid support is a aminoalkyl moiety.
Claim 4
Dependent Claim 24 words
A probe according to claim 1, characterized in that the chemical linkage to the solid support is a : Hg-S or a S-Hg moiety.
Claim 5
Dependent Claim 20 words
A probe according to claim 1, characterized in that the chemical linkage to the solid support is a -S-S- moiety.
Claim 6
Dependent Claim 25 words
A probe according to claims 1 - 4, characterized in that the nucleic acid is a ribonucleic acid or a deoxyribonucleic acid or an oligonucleotide.
Claim 7
Dependent Claim 14 words
A probe having a nucleic acid linked thereto of the formula nucleic acid-CH=N-solid support.
Claim 8
Dependent Claim 14 words
A probe having a nucleic acid linked thereto of the formula nucleic acid-CH=N-solid support.
Claim 9
Dependent Claim 7 words
A nucleic acid-containing material of the formula
Claim 10
Dependent Claim 7 words
A nucleic acid-containing material of the formula
Claim 11
Dependent Claim 15 words
A nucleic acid-nucleotide unit including a purine or pyrimidine containing a Hg or -SH moiety.
Claim 12
Dependent Claim 15 words
A nucleic acid-nucleotide unit including a purine or pyrimidine containing a Hg or -SH moiety.
Claim 13
Dependent Claim 95 words
A process for producing a probe immobilized to a solid support, characterized in that a ribonucleic acid or a oligoribonucleotide or a deoxyribonucleic acid or a deoxyoligoribonucleotide, said deoxyribonucleic acid or deoxyoligoribonucleotide being bound to a substituted sugar moiety, containing vicinale hydroxy- groups, is reacted with periodate thereby to form a nucleic acid having a pendent aldehyde group, reacting the product with a solid support having a pendent primary amino group thereby to form a Schiff's base of the formula nucleic acid -CH=N-solid support and reducing said Schiff's base thereby to form the desired probe.
Claim 14
Dependent Claim 95 words
A process for producing a probe immobilized to a solid support, characterized in that a ribonucleic acid or a oligoribonucleotide or a deoxyribonucleic acid or a deoxyoligoribonucleotide, said deoxyribonucleic acid or deoxyoligoribonucleotide being bound to a substituted sugar moiety, containing vicinale hydroxy- groups, is reacted with periodate thereby to form a nucleic acid having a pendent aldehyde group, reacting the product with a solid support having a pendent primary amino group thereby to form a Schiff's base of the formula nucleic acid -CH=N-solid support and reducing said Schiff's base thereby to form the desired probe.
Claim 15
Dependent Claim 97 words
A process for producing a probe immobilized to a solid support, characterized in that a ribonucleic acid or a deoxyribonucleic acid or an oligonucleotide is attached to a modified ribonucleotide or deoxynucleotide carrying on its purine or pyrimide ring a (a) Hg or (b) SH radical and reacting the coupled nucleic acid- modified nucleotide with a solid support having a penden group, in case of (a) a SH and in case of (b) a SH or Hg thereby to form the desired probe, linked to the said solid support with a Hg-S or S-Hg or -S-S- moiety.
Claim 16
Dependent Claim 97 words
A process for producing a probe immobilized to a solid support, characterized in that a ribonucleic acid or a deoxyribonucleic acid or an oligonucleotide is attached to a modified ribonucleotide or deoxynucleotide carrying on its purine or pyrimide ring a (a) Hg or (b) SH radical and reacting the coupled nucleic acid- modified nucleotide with a solid support having a penden group, in case of (a) a SH and in case of (b) a SH or Hg thereby to form the desired probe, linked to the said solid support with a Hg-S or S-Hg or -S-S- moiety.
Claim 17
Independent Claim 72 words
A method for determining whether a ribonuclej acid or a deoxynucleic acid or an oligonucleotide contai ned in a liquid test sample includes a particular nucle: acid sequence wherein the liquid sample is contacted with a separation probe comprising a nucleic acid immobilized to a solid support by a single chemical linkage and the separation probe and/or the remaining liquid sample is subjected to analysis to ascertain any change in its composition.
Claim 18
Independent Claim 72 words
A method for determining whether a ribonuclej acid or a deoxynucleic acid or an oligonucleotide contai ned in a liquid test sample includes a particular nucle: acid sequence wherein the liquid sample is contacted with a separation probe comprising a nucleic acid immobilized to a solid support by a single chemical linkage and the separation probe and/or the remaining liquid sample is subjected to analysis to ascertain any change in its composition.

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