End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids

• US 5,237,016 A
• Filed: 01/05/1989
• Issued: 08/17/1993
• Est. Priority Date: 01/05/1989
• Status: Expired due to Fees

• Current Assignee: Akzo Nobel N.V. (Nouryon BV)
• Original Assignee: Siska Diagnostics, Inc.
• Inventors: Fahy, Eoin D., Ghosh, Soumitra S.
• Primary Examiner(s): Brown, Johnnie R.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US07/293,893
• Time in Patent Office: 1,685 Days
• Field of Search: 536/29, 536/27, 935/78, 435/6, 435/91, 435/810, 435/803, 436/501, 528/332, 526/310, 525/329.4
• US Class Current: 525/329.4
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

This invention concerns methods and means for covalent attachment of oligonucleotides to solid supports substantially at their 5'"'"'-ends. According to the invention thiol-oligonucleotides are attached to bromoacetyl-derivatized polyacrylamide supports, or conversely, bromoacetyl-oligonucleotides are immobilized on thiol-polyacrylamide supports.

In a further aspect, this invention relates to bromoacetyl-oligonucleotides that may be immobilized on thiol-polyacrylamide solid supports, thiol-oligonucleotides immobilized on bromoacetyl-derivatized polyacrylamide supports as well as to methods for capture of nucleic acids by oligonucleotides attached to polyacrylamide solid supports, either by direct capture or in sandwich hybridization formats.

Claims

1. Polyacrylamide supports to which RNA or DNA oligonucleotides are covalently attached via a thioether linkage, wherein:

at least about 95% of the oligonucleotides are linked to the carboxamide side groups of the support at the 5'"'"'-ends of the oligonucleotides;

the polyacrylamide supports have a molecular weight exclusion limit of at least about 2×

10⁷ daltons; and

the supports to which the oligonucleotides are covalently attached via a thioether linkage are selected from the group consisting of supports represented by the formulae;

##STR1##

2. The supports of claim 1 wherein the repeating units each contain three hydroxymethyl groups and one secondary amide group.

3. The supports of claim 1 wherein the oligonucleotides are about 12 to 200 nucleotides in length.

4. The supports of claim 1 wherein the oligonucleotides are about 15-40 nucleotides in length.

5. Oligonucleotides derivatized at their 5'"'"'-terminal phosphate group with bromoacetyl group and having the following formula:

##STR2##