Coupling of nucleic acids to solid support by photochemical methods

• US 4,713,326 A
• Filed: 05/18/1984
• Issued: 12/15/1987
• Est. Priority Date: 07/05/1983
• Status: Expired due to Term

• Current Assignee: Miles, Inc. (Bayer AG)
• Original Assignee: Molecular Diagnostics, Inc. (Medite Cancer Diagnostics, Inc.)
• Inventors: Crothers, Donald M., Dattagupta, Nanibhushan
• Primary Examiner(s): NOT, DEFINED
• Application Number:
  US06/611,667
• Time in Patent Office: 1,306 Days
• Field of Search: 435/6, 422/57, 436/63, 436/94, 436/501, 935/78
• US Class Current: 435/6.12
• 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: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: G01N 33/548 Carbohydrates, e.g. dextran
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A solid support capable of binding a nucleic acid thereto upon suitable irradiation, comprising (a) a solid substrate, (b) a photochemically reactive intercalator compound or other nucleic acid-binding ligands, and (c) divalent radical chemically linking the substrate and the ligand (b). Specifically, a hydroxy group-containing solid substrate such as nitrocellulose paper is linked via a bifunctional reagent such as cyanogen bromide or 1,4-butanedioldiglycidyl ether to an amino-substituted angelicin or psoralen or ethidium bromide which in turn is photochemically linked to a nucleic acid. The resulting immobilized nucleic acid probe is capable of hybridizing with complementary nucleic acid fragments and is thereby useful in diagnostic assays.

Claims

1. A solid support capable of binding a nucleic acid thereto upon suitable irradiation, comprising (a) a solid substrate, (b) a photochemically reactive nucleic acid-binding ligand, and (c) a divalent radical chemically linking the substrate and the nucleic acid-binding ligand.

2. A support according to claim 1, wherein the nucleic acid-binding ligand is an intercalator compound selected from the group consisting of acridine dyes, phenazines, phenothiazines, and quinolines.

3. A support according to claim 1, wherein the substrate (a) in free state has free OH groups through which it is linked by the divalent radical (c).

4. A support according to claim 1, wherein the substrate (a) is cellulose or a cellulose ester.

5. A support according to claim 1, wherein the divalent radical (c) is derived from cyanogen bromide.

6. A support according to claim 1, wherein the divalent radical (c) is derived from 1,4-butanediol-diglycidyl ether.

7. An immobilized nucleic acid probe comprising (a) a nucleic acid, (b) a nucleic acid-binding ligand photochemically linked to the nucleic acid, and (c) a solid substrate chemically linked through a divalent radical to the nucleic acid-binding ligand (b).

8. An immobilized probe according to claim 7, wherein the nucleic acid-binding ligand is an intercalator compound selected from the group consisting of acridine dyes, phenazines, phenothiazines, and quinolines.

9. An immobilized probe according to claim 7, wherein the substrate (c) in free state has free OH groups through which it is linked by the divalent radical.

10. An immobilized probe according to claim 7, wherein the substrate (c) is cellulose or a cellulose ester.

11. An immobilized probe according to claim 7, wherein the divalent radical is derived from cyanogen bromide.

12. An immobilized probe according to claim 7, wherein the divalent radical is derived from 1,4-butanediol-diglycidyl ether.