UV excitable fluorescent energy transfer dyes

• US 8,129,529 B2
• Filed: 12/02/2010
• Issued: 03/06/2012
• Est. Priority Date: 08/27/1999
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Lee, Linda G.
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US12/959,205
• Publication Number:
  US 20110077415A1
• Time in Patent Office: 460 Days
• Field of Search: 546/1, 546/16, 546/34, 548/219, 435/6, 536/26.6
• US Class Current: 546/1
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Novel energy transfer dyes which can be used with shorter wavelength light sources are provided. These dyes include a donor dye with an absorption maxima at a wavelength between about 250 to 450 nm and an acceptor dye which is capable of absorbing energy emitted from the donor dye. One of the energy transfer dyes has a donor dye which is a member of a class of dyes having a coumarin or pyrene ring structure and an acceptor dye which is capable of absorbing energy emitted from the donor dye, wherein the donor dye has an absorption maxima between about 250 and 450 nm and the acceptor dye has an emission maxima at a wavelength greater than about 500 nm.

Claims

1. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 300 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima greater than about 500 nm.

2. The energy transfer dye according to claim 1, wherein the donor dye has an absorption maxima at a wavelength between about 350 and 400 nm.

3. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 250 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima at a wavelength greater than about 550 nm.

4. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 250 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima at a wavelength between about 500 and 700 nm.

5. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 250 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima at a wavelength at least about 150 nm greater than the absorption maxima of the donor dye.

6. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 300 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima greater than about 500 nm,

wherein the acceptor dye is a member of a class of dyes selected from the group consisting of fluorescein, rhodamine, asymmetric benzoxanthene, xanthene, cyanine, phthalocyanine and squaraine dyes.

7. The energy transfer dye according to claim 6, wherein the acceptor dye is selected from the group consisting of 4,7-dichlorofluorescein dyes, asymmetric benzoxanthene dyes, rhodamine, 4,7-dichlororhodamine dyes, carboxyrhodamines, N,N,N′

,N′

-tetramethyl carboxyrhodamines, carboxy R110, carboxy R6G, carboxy-X-rhodamines and Cy5.

8. The energy transfer dye according to claim 6, wherein the acceptor dye is selected from the group consisting of R110, R6G, TAMRA and ROX.

9. An energy transfer dye, comprising:

(a) a donor dye having an absorption maxima at a wavelength between about 300 and about 450 nm; and

(b) an acceptor dye capable of absorbing excitation energy emitted by the donor dye and emitting light in response, the acceptor dye having an emission maxima greater than about 500 nm,

further comprising a linker attaching the donor dye to the acceptor dye.

10. The energy transfer dye according to claim 9, wherein the linker linking the donor dye to the acceptor dye is such that the acceptor dye absorbs substantially all of the excitation energy emitted by the donor dye, the linker including a functional group selected from the group consisting of an alkene, diene, alkyne, a five-membered ring and a six-membered ring, each of the five-member ring and the six-member ring having at least one unsaturated bond or a fused ring structure.

11. The energy transfer dye according to claim 10, wherein the linker functional group is a five- or six-membered ring selected from the group consisting of cyclopentene, cyclohexene, cyclopentadiene, cyclohexadiene, furan, thiofuran, pyrrole, isopyrrole, isoazole, pyrazole, isoimidazole, pyran, pyrone, benzene, pyridine, pyridazine, pyrimidine, pyrazine oxazine, indene, benzofuran, thionaphthene, indole and naphthalene.