Patent EA-003929-B1 (EA)
Title: Nucleic acid modified oligomer, method of producing same, modified conductive surface, method of producing same and method for electrochemically detecting nucleic acid-oligomer hybridisation events
Inventor: HARTWICH GERHARD
IPC: C12N15/09
Filed: 2000-01-07
Abstract: 1. A nucleic acid oligomer modified by covalently attaching a redox-active moiety, characterized in that the redox-active moiety comprises at least one electron-donor molecule and at least one electron-acceptor molecule, the electron-donor molecule and electron-acceptor molecule not being joined with one another by nucleic acid oligomers. 2. The modified nucleic acid oligomer according to claim 1, characterized in that the redox-active moiety comprises at least one redox-active, linked, at least bimolecular electron-donor/electron-acceptor complex, at least one electron-donor molecule of the redox-active moiety and at least one electron-acceptor molecule of the redox-active moiety being joined with one another via one or more bonds. 3. The modified nucleic acid oligomer according to claim 2, characterized in that the bonds are covalent bonds. 4. The modified nucleic acid oligomer according to claim 1, characterized in that the redox-active moiety comprises at least one redox-active, linked, at least bimolecular electron-donor/electron-acceptor complex, at least one electron-donor molecule of the redox-active moiety and at least one electron-acceptor molecule of the redox-active moiety being covalently joined via one or more branched or linear molecular moieties of any composition and chain length. 5. The modified nucleic acid oligomer according to claim 4, characterized in that the branched or linear molecular moieties have a chain length of 1 - 20 atoms. 6. The modified nucleic acid oligomer according to claim 5, characterized in that the branched or linear molecular moieties have a chain length of 1 - 14 atoms. 7. The modified nucleic acid oligomer according to one of the preceding claims, characterized in that the redox-active moiety additionally comprises one or more macromolecules. 8. The modified nucleic acid oligomer according to one of the preceding claims, wherein the redox-active moiety is the native or modified reaction center of photosynthesizing organisms. 9. The modified nucleic acid oligomer according to claim 8, wherein the redox-active moiety is the native or modified reaction center of photosynthesizing bacteria. 10. The modified nucleic acid oligomer according to one of claims 1 through 7, characterized in that at least one of the electron-donor molecules and electron-acceptor molecules is a pigment. 11. The modified nucleic acid oligomer according to claim 10, characterized in that the pigment is a flavin, a (metallo)porphyrin, a (metallo)chlorophyll, a (metallo)bacteriochlorophyll, or a derivative of these pigments. 12. The modified nucleic acid oligomer according to one of claims 1 through 7, characterized in that at least one of the electron donor molecules and electron acceptor molecules is a nicotinamide or a quinone. 13. The modified nucleic acid oligomer according to claim 12, characterized in that the quinone is a pyrroloquinoline quinone (PQQ), a 1,2-benzoquinone, a 1,4-benzoquinone, a 1,2-naphthoquinone, a 1,4-naphthoquinone, a 9,10-anthraquinone, or one of their derivatives. 14. The modified nucleic acid oligomer according to one of claims 1 through 7, characterized in that at least one of the electron-donor molecules and electron-acceptor molecules is a charge transfer complex. 15. The modified nucleic acid oligomer according to claim 14, characterized in that the charge transfer complex is a transition metal complex. 16. The modified nucleic acid oligomer according to claim 15, characterized in that the charge transfer complex is a Ru(II), Cr(III), Fe(II), Os(II), or Co(II) complex. 17. The modified nucleic acid oligomer according to one of the preceding claims, characterized in that the modified nucleic acid oligomer can sequence-specifically bind single-strand DNA, RNA, and/or PNA. 18. The modified nucleic acid oligomer according to claim 17, characterized in that the modified nucleic acid oligomer is a deoxyribonucleic acid oligomer, a ribonucleic acid oligomer, or a peptide nucleic acid oligomer. 19. The modified nucleic acid oligomer according to one of the preceding claims, characterized in that the redox-active moiety is covalently bound to one of the phosphoric-acid groups, to one of the carboxylic-acid groups, to one of the amine groups, or to a sugar of the nucleic acid oligomer backbone. 20. The modified nucleic acid oligomer according to one of the preceding claims, characterized in that the redox-active moiety is covalently bound to a sugar-hydroxy group of the nucleic acid oligomer backbone. 21. The modified nucleic acid oligomer according to one of claims 1 through 18, characterized in that the redox-active moiety is covalently attached to a thiol group, a hydroxyl group, a carboxylic-acid group, or an amine group of a modified base of the nucleic acid oligomer. 22. The modified nucleic acid oligomer according to claim 21, characterized in that the reactive thiol, hydroxyl, carboxylic-acid, or amine group of the base is covalently bound to the base via a branched or linear molecular moiety of any composition and chain length, the shortest continuous link between the thiol, hydroxyl, carboxylic-acid, or amine group and the base being a branched or linear molecular moiety having a chain length of 1 - 20 atoms. 23. The modified nucleic acid oligomer according to claim 22, characterized in that the shortest continuous link between the thiol, hydroxyl, carboxylic-acid, or amine group and the base is a branched or linear molecular moiety having a chain length of 1 - 14 atoms. 24. The modified nucleic acid oligomer according to one of claims 19 through 22, characterized in that the redox-active moiety is attached to an end of the nucleic acid oligomer backbone or to a terminal modified base. 25. The modified nucleic acid oligomer according to one of the preceding claims, characterized in that the redox-active moiety is a photoinducibly redox-active moiety. 26. The modified nucleic acid oligomer according to one of claims 1 through 24, characterized in that the redox-active moiety is a chemically-inducibly redox-active moiety. 27. A method of producing a modified nucleic acid oligomer as defined in one of the preceding claims, characterized in that a redox-active moiety is covalently attached to a nucleic acid oligomer moiety by one or more amidations with amine or acid groups of the redox-active moiety by one or more esterifications with alcohol or acid groups of the redox-active moiety, by thioester formation with thioalcohol or acid groups of the redox-active moiety, or by condensation of one or more amine groups of the nucleic acid oligomer with aldehyde groups of the redox-active moiety and subsequent reduction of the resultant carbon-nitrogen double bond. 28. The method of producing a modified nucleic acid oligomer according to claim 27, characterized in that the redox-active moiety is attached to a nucleic acid oligomer by covalently attaching at least one electron-donor molecule. 29. The method of producing a modified nucleic acid oligomer according to claim 27, characterized in that the redox-active moiety is attached to a nucleic acid oligomer by covalently attaching at least one electron-acceptor molecule. 30. The method of producing a modified nucleic acid oligomer according to claim 27, characterized in that the redox-active moiety is attached to a nucleic acid oligomer by covalently attaching at least one macromolecule or by covalently attaching at least one protein. 31. The method of producing a modified nucleic acid oligomer according to claims 28 through 30, characterized in that the redox-active moiety is completed by adding at least one component selected from the group consisting of electron-acceptor molecules, electron-donor molecules, macromolecules, and proteins. 32. The method of producing a modified nucleic acid oligomer according to one of claims 27 through 31, characterized in that at least one branched or linear molecular moiety of any composition and chain length is covalently attached to the redox-active moiety and the branched or linear molecular moiety has a reactive amine, hydroxyl, thiol, acid, or aldehyde group for covalent attachment to a nucleic acid oligomer. 33. The method of producing a modified nucleic acid oligomer according to claim 32, characterized in that the shortest continuous link between the nucleic acid oligomer and the redox-active moiety is a branched or linear molecular moiety having a chain length of 1 - 20 atoms. 34. The method of producing a modified nucleic acid oligomer according to claim 33, characterized in that the shortest continuous link between the nucleic acid oligomer and the redox-active moiety is a branched or linear molecular moiety having a chain length of 1 - 14 atoms. 35. A modified conductive surface, characterized in that at least one type of modified nucleic acid oligomer according to one of claims 1 through 26 is attached to a conductive surface. 36. The modified conductive surface according to claim 35, characterized in that the surface consists of a metal or a metal alloy. 37. The modified conductive surface according to claim 36, characterized in that the surface consists of a metal selected from the group: platinum, palladium, gold, cadmium, mercury, nickel, zinc, carbon, silver, copper, iron, lead, aluminum, and manganese. 38. The modified conductive surface according to claim 35, characterized in that the surface consists of a semiconductor. 39. The modified conductive surface according to claim 38, characterized in that the surface consists of a semiconductor selected from the group: carbon, silicon, germanium, and tin. 40. The modified conductive surface according to claim 35, characterized in that the surface consists of a binary compound of the elements of groups 14 and 16, a binary compound of the elements of groups 13 and 15, a binary compound of the elements of groups 15 and 16, or a binary compound of the elements of groups 11 and 17. 41. The modified conductive surface according