Photochemical nucleic acid-labeling reagent having a polyalklamine spacer

• US 5,026,840 A
• Filed: 02/06/1990
• Issued: 06/25/1991
• Est. Priority Date: 01/10/1985
• Status: Expired due to Fees

• Current Assignee: Miles, Inc. (Bayer AG)
• Original Assignee: Molecular Diagnostics, Inc. (Medite Cancer Diagnostics, Inc.)
• Inventors: Dattagupta, Nanibhushan, Albarella, James P.
• Primary Examiner(s): Rollins, John W.
• Application Number:
  US07/475,639
• Time in Patent Office: 504 Days
• Field of Search: 536/27, 435/4, 435/6, 435/188, 436/63, 436/94, 436/501
• US Class Current: 536/25.32
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A photochemical nucleic acid-labeling reagent of the formula ##STR1## wherein Q is a photoreactive residue of a nucleic acid-binding ligand; L is a detectable label residue; R is hydrogen, C₁ to C₇ -alkyl, aryl, hydroxy, or C₁ to C₇ -alkoxy; x is an integer from 2 through 7; and Y is an integer from 3 through 10; wherein R and x, respectively, can be the same or different each time they appear in the formula. The reagent is useful in the highly efficient labeling of nucleic acids for the purpose of detection in hybridization assays.

Claims

1. A photochemical nucleic acid-labeling reagent of the formula ##STR11## wherein Q is a photoreactive residue of a nucleic acid-binding ligand;

L is a detectable label residue;

R is hydrogen, C₁ to C₇ -alkyl, aryl, hydroxy, or C₁ to C₇ -alkoxy;

wherein R can be the same or different each time R appears in the formula.

2. A reagent according to claim 1, wherein R is hydrogen or C₁ to C₄ -alkyl.

3. A reagent according to claim 1, wherein the photoreactive residue of a nucleic acid-binding ligand is a photoreactive residue of an intercalator compound.

4. A reagent according to claim 3, wherein the intercalator compound is selected from the group consisting of acridine dyes, phenanthridines, phenazines, furocoumarins, phenothiazines, quinolines and antracyclines.

5. A reagent according to claim 4, wherein the intercalator compound is selected from the group consisting of furocoumarins and phenanthridines.

6. A reagent according to claim 3, wherein the intercalator compound is of the formula ##STR12## in which R¹, R² and R³ each independently is hydrogen or C₁ -C₇ -alkyl;

and R⁴ is hydrogen, C₁ -C₇ -alkyl or C₁ -C₇ -alkyl substituted with hydroxy, C₁ -C₇ -alkoxy, amino, halo, or is of the formula ##STR13##

7. A reagent according to claim 3, wherein the intercalator compound is 4'"'"'-aminomethyl-4,5'"'"'-dimethylangelicin.

8. A reagent according to claim 3, wherein the intercalator compound is of the formula ##STR14## in which R¹, R³ and R⁶ each independently is hydrogen or C₁ -C₇ -alkyl;

R⁴ is hydrogen, C₁ -C₇ -alkyl or C₁ -C₇ -alkyl substituted with hydroxy, C₁ -C₇ -alkoxy, amino, halo, or is of the formula ##STR15## R² and R⁵ each independently is hydrogen, hydroxy, carboxy, carbo-C₁ -C₇ -alkoxy or C₁ -C₇ -alkoxy.

9. A reagent according to claim 3, wherein the intercalator compound is 4'"'"'-aminomethyl-4,5'"'"',8-trimethylpsoralen.

10. A reagent according to claim 1, wherein the detectable label residue is a residue of a moiety selected from the group consisting of biotin, a hapten, an enzyme, a fluorescent molecule and a luminescent molecule.

11. A reagent according to claim 1, wherein the detectable label residue is selected from the group consisting of a biotin and a hapten residue.

12. A labeled nucleic acid comprising (a) a nucleic acid component, (b) a nucleic acid-binding ligand photochemically linked to the nucleic acid component, (c) a label, and (d) a spacer chemically linking (b) and (c) and being of the formula ##STR16## wherein R is hydrogen, C₁ -C₇ alkyl, aryl, hydroxy, or C₁ -C₇ alkoxy;

wherein R can be the same or different each time R appears in the formula.

13. A labeled nucleic acid according to claim 12, wherein R is hydrogen or C₁ -C₄ alkyl.

14. A labeled nucleic acid according to claim 12, wherein the nucleic acid-binding ligand is an intercalator compound.

15. A labeled nucleic acid according to claim 14, wherein the intercalator compound is selected from the group consisting of acridine dyes, phenanthridines, phenazines, furocoumarins, phenothiazines, quinolines and anthracyclines.

16. A labeled nucleic acid according to claim 15, wherein the intercalator compound is selected from the group consisting of furocoumarins and phenanthridines.

17. A labeled nucleic acid according to claim 14, wherein the intercalator compound is of the formula ##STR17## in which R¹, R² and R³ each independently is hydrogen or lower alkyl;

and R⁴ is hydrogen, C₁ -C₇ -alkyl or C₁ -C₇ -alkyl substituted with hydroxy, C₁ -C₇ -alkoxy, amino, halo, or is of the formula ##STR18##

18. A labeled nucleic acid according to claim 14, wherein the intercalator compound is 4'"'"'-aminomethyl-4,5'"'"'-dimethylangelicin.

19. A labeled nucleic acid according to claim 14, wherein the intercalator compound is of the formula ##STR19## in which R¹, R³ and R⁶ each independently is hydrogen or C₁ -C₇ -alkyl;

R⁴ is hydrogen, C₁ -C₇ -alkyl or C₁ -C₇ -alkyl substituted with hydroxy, C₁ -C₇ -alkoxy, amino, halo, or is of the formula ##STR20## R² and R⁵ each independently is hydrogen, hydroxy, carboxy, carbo-C₁ -C₇ -alkoxy or C₁ -C₇ -alkoxy.

20. A labeled nucleic acid according to claim 14, wherein the intercalator compound is 4'"'"'-aminomethyl-4,5'"'"'-8-trimethylpsoralen.

21. A labeled nucleic acid according to claim 12, wherein the label is selected from the group consisting of biotin, a hapten, an enzyme, a fluorescent molecule and a luminescent molecule.

22. A labeled nucleic acid according to claim 12, wherein the label is selected from the group consisting of biotin and a hapten.

23. A labeled nucleic acid according to claim 12, wherein the nucleic acid component is DNA.

24. A labeled nucleic acid according to claim 12, wherein the nucleic acid component is RNA.

25. A labeled nucleic acid according to claim 12, wherein the nucleic acid component is an oligonucleotide.