| 摘要 |
<p>#CMT# #/CMT# Use of at least two different fluorophores for configuring a fluorescence resonance energy transfer pair (FRET pair), is claimed, where: at least one first fluorophore (A) serves as the donor fluorophore and at least one second fluorophore (B) serves as the acceptor fluorophore within the FRET pair, the first fluorophore (A) and the second fluorophore (B), independently of each other, each are configured as the basis of an organo-metal complex of rare earth element, and the fluorophores (A) and (B) comprise different rare earth elements from each other. #CMT# : #/CMT# Independent claims are included for: (1) FRET complex comprising at least a first fluorophore (A) as a donor fluorophore and at least a second fluorophore (B) as an acceptor fluorophore, where: the fluorophore (A) and (B) are coupled and/or bonded over a bivalent and/or double bonded organic residue, preferably a linker and/or spacer, the first fluorophore (A) and the second fluorophore (B) forms an organometal complexes of rare earth elements and the fluorophore (A) and (B) contains different rare earth elements; (2) a FRET system comprising at least one first fluorophore (A) as a donor fluorophore and at least one second fluorophore (B) as an acceptor fluorophore; (3) a use of the FRET complexes or FRET system for the detection of an event in a sample; and (4) a FRET pair comprising at least two different fluorophores, where the FRET pair comprises the first fluorophore (A) as a donor fluorophore and the second fluorophore (B) as an acceptor fluorophore. #CMT#USE : #/CMT# The two different fluorophores are useful for configuring FRET pair. The FRET complexes or system is useful for: the detection of an event in a sample; the interaction with the target molecule and/or structure, and the detection and/or identification of the target molecule and/or structure, preferably nucleotide sequence, preferably DNA or RNA sequence, where the structure is a (poly)peptide and/or an amino acid sequence, preferably an enzyme or an antibody. The FRET complexes or system is useful: as a dye, which is a fluorescence dye, preferably useful for marking purposes; and in or as biosensors, probes, preferably FRET-probes (all claimed). #CMT#ADVANTAGE : #/CMT# The fluorophores provides defined emission spectrum with a very narrow emission band for long time, which results in an excellent discrimination of the desired measured signal, when compared to the non-specific radiation. #CMT#INORGANIC CHEMISTRY : #/CMT# Preferred Components: The first fluorophore (A) and the second fluorophore (B) are selected in such a way that under the influence, effect and/or entry of stimulation energy, an energy transfer of the first fluorophore (A) to the second fluorophore (B) takes place, where the energy transfer of the first fluorophore (A) to the second fluorophore (B) preferably takes place in radiation free manner. The energy transfer of the first fluorophore (A) to the second fluorophore (B) is dependent on the spacing of the fluorophore (A) and (B), where the energy transfer, preferably the intensity and/or effect of the energy transfer increases with decrease in the spatial spacing of the fluorophore (A) and (B). The energy transfer of the first fluorophore (A) to the second fluorophore (B) leads to the removal of fluorescence of the first fluorophores (A) and to increase the fluorescence of the second fluorophore (B). The first fluorophore (A) partially overlaps with the second fluorophore (B). The first fluorophore (A) and/or second fluorophore (B) comprise a dye, preferably a fluorescence dye. The first fluorophore (A) and/or the second fluorophore (B) exhibit at least a core, preferably a core from rare earth element. The rare earth element is cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium (all preferred), scandium, yttrium or lanthanum. The fluorophore (A) is terbium, preferably in the form of terbium(III). The fluorophore (B) is europium, preferably in the form of europium(III). The rare earth elements in the fluorophores (A) and (B) are respectively complexed with at least a ligand, preferably complex forming agent and/or chelate forming agent, where: the ligands are polyvalent ligands, preferably tetravalent ligands, and the rare earth elements in the fluorophore (A) and (B) are bonded to the ligands in an ionic-, coordinative- and/or covalent-manner (preferred). The ligand is a polydentate (preferably bidentate) ligand, which is picolinic acid, picolinate and/or its derivative, preferably hydroxy derivatives; or picolinic acid, picolinate and/or its derivative, substituents and/or functional groups exhibiting covalent bond and/or couplings to organic molecules, where the substituents and/or functional groups are amino-, carboxylate-, isocyanate-, thioisocyanate-, epoxy-, thiol- or hydroxyl- group (preferred). The ligand is preferably hydroxy picolinic acid and/or hydroxy picolinate. The fluorophore (A) is a terbium containing complex of formula (V), preferably tetra(4-hydroxypyridin-2-carboxylato)terbium(III), tris(pyridin-2-carboxylato)-(4-hydroxypyridin-2-carboxylato)terbium(III), bis(pyridin-2-carboxylato)-bis(4-hydroxypyridin-2-carboxylato)terbium(III), (pyridin-2-carboxylato)-tris(4-hydroxypyridin-2-carboxylato)terbium(III) and/or its derivative. The fluorophore (A) is of formula ((Tb x(Pic) y(Pic-Y 1>) z)(4-3x) ->) (I). The fluorophore (B) is of formula ((Eu x(Pic) y(Pic-Y 1>) z)(4-3x) ->) (I). The fluorophore (B) is europium containing complex of formula (IV), preferably tetra (4-hydroxypyridin-2-carboxylato)europium(III), tris(pyridin-2-carboxylato)-(4-hydroxypyridin-2-carboxylato)europium(III), bis(pyridin-2-carboxylato)-bis(4-hydroxypyridin-2-carboxylato)europium(III), (pyridin-2-carboxylato)-tris(4-hydroxypyridin-2-carboxylato)europium(III) and/or its derivative. The organic residue is selected in such a manner that between the fluorophore (A) and fluorophore (B), a fluorescence energy transfer takes place. The organic residue is covalently bonded with the fluorophore (A) and/or the fluorophore (B), where the organic residue is covalently bonded and/or coupled with the functional groups of the fluorophore (A) and/or (B), preferably the organic residues covalently binds and/or couples with the functional groups or ligands exhibiting the functional groups. The organic residue is covalently bonded with the fluorophore (A) and/or fluorophore (B) to form (poly) urethane group. The interaction of the organic residue with the target molecule and/or target structure leads to a change, preferably a reduction in the FRET of the fluorophore (A) to the fluorophore (B). The fluorophore (A) and fluorophore (B) of the FRET complex and/or system in the sample forms a FRET pair. The fluorophore (A) and fluorophore (B) with an organic residue is a molecule of formula (III). The FRET complex is of formula (DF 1>-S 1>-AF 1>). The FRET complex and/or the FRET system during the entry of the event and/or as a result of the event preferably under the influence, effect and/or entry of a stimulation energy forms: a mutation in its emission spectrum and/or a detectable signal. The change of the emission spectrum and/or detectable signal retards in a temporal sequence with respect to the entry of the event, and/or the change of the emission spectrum and/or detectable signal takes place in temporal sequence after the entry of the event. The detection of the change in the emission spectrum and/or detectable signal takes place through a time scattering of fluorescence measurement, preferably through the autofluorescence of the sample. The detectable signal of the FRET complex and/or system in comparison with the autofluorescence of the sample is preferably a factor of 2, preferably 10. The event is an interaction and/or hybridization of the FRET complex and/or FRET system with a target molecule and/or a target structure, preferably the event is a spatial change, which is a configurational or conformational change of the FRET complexes and/or FRET system. DF 1>donorfluorophore, preferably fluorophore (A); AF 1>acceptor fluorophore, preferably fluorophore (B); S 1>preferably a divalent and/or double bonded organic residue, preferably a linker and/or spacer; n : 1.5-20, preferably 2-12; Tb : terbium(III); Pic : picolinate; Y 1>a functional group, preferably amino-, carboxylate-, isocyanate-, thioisocyanate-, epoxy-, thiol- or OH (preferred); Eu : europium(III); x : 1-4, preferably 1; and either y, z : 0-4; or y+z : 4. #CMT#[Image]#/CMT# #CMT#[Image]#/CMT# #CMT#[Image]#/CMT# #CMT#ORGANIC CHEMISTRY : #/CMT# Preferred Components: The organic residue: is a residue, with which a target molecule and/or a target structure is capable of interaction, and/or is a residue, which is an integratable with the target molecule and/or target structure. The organic residue is a nucleotide sequence, which is a DNA- or RNA- sequence, preferably an oligonucleotide. The nucleotide sequence, preferably exhibits a complementary nucleotide sequence segments at its 5'-terminal and 3'-terminal, such that the non interaction with the target molecule and/or target structure results in hybridization of the complementary nucleotide sequence segment. The fluorophore (A) and fluorophore (B) forms a molecular beacon by covalently bonding the nucleotide sequence with 5'-terminal and 3'-terminal complementary nucleotide sequence segments. The organic residue is a peptide and/or an amino acid sequence, preferably an oligopeptide; an optionally saturated, preferably unsaturated hydrocarbyl residue. The chain length of the hydrocarbyl residue is at least 2 carbon atoms, preferably 2-12 carbon atoms. #CMT#EXAMPLE : #/CMT# No suitable example given.</p> |
