Oligonucleotide-polyamide conjugates

• US 5,677,440 A
• Filed: 02/09/1996
• Issued: 10/14/1997
• Est. Priority Date: 07/16/1990
• Status: Expired due to Term

• Current Assignee: Howard Florey Institute of Experimental Physiology and Medicine
• Original Assignee: Howard Florey Institute of Experimental Physiology and Medicine
• Inventors: Tregear, Geoffrey William, Haralambidis, Jim
• Primary Examiner(s): Marschel, Ardin H.
• Application Number:
  US08/599,193
• Time in Patent Office: 613 Days
• Field of Search: 435/6, 435/810, 436/501, 536/23.1, 536/24.1, 536/25.3, 935/78
• US Class Current: 536/25.3
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• 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
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Conjugates of the type Z--X--L are described wherein Z is a solid phase matrix, X is a single amino acid or a polyamide of two or more amino acid residues, and L is a linker that is capable of linking between the amino terminus of X and a 3'"'"'-phosphate of a nucleotide. Conjugates may also be synthesized by assembling a polyamide on a solid support matrix, adding a suitable linker molecule, followed by synthesis of an oligonucleotide thereon. Methods for detecting specific polynucleotides with oligonucleotide-polyamide conjugates are also described.

Claims

1. A composition having the formula Z--X--L, where Z represents a solid phase matrix, X represents a single amine acid or a polyamide comprising two or more amine acid residues linked through its C-terminus to said solid phase matrix, and L represents a linker attached to the N-terminus of the amine acid or polyamide wherein said linker has a reactive group which is capable of forming a bond with a 3'"'"' phosphate of a nucleotide, and said linker is derived from a) an α

,ω

hydroxy carboxylic acid derivative, b) a lactone, or c) a bifunctional linker having the formula R'"'"'--L'"'"'--R", wherein L'"'"' is an alkyl optionally substituted with one or more halogen, cyano, phenyl, amino, carboxy, alkyl, or alkoxy, and optionally interrupted by one or more of --S--, --O--, --NH--, --CH═

CH--, --C═

C--, phenyl or --SO₂ --, R'"'"' is a functional group that covalently binds to the N-terminus of said polyamide, and R" is a functional group that is the same as or different from R'"'"' that is capable of covalently binding to the 3'"'"' phosphate group of a nucleotide.

2. A composition according to claim 1, wherein said single amino acid or polyamide X contains one or more reporter groups.

3. A composition according to claim 2 wherein said reporter group is selected from the group consisting of fluorophores, biotin, enzymes, and colloidal compounds.

4. A composition according to claim 2 wherein said reporter group is selected from the group consisting of fluorescein, tetramethyl rhodamine, Texas Red, coumarins, carbonic anyhdrase, urease, horseradish peroxidase, dehydrogenases, colloidal gold, and colloidal silver.