Reagent for detecting an analyte

• US 20110111519A1
• Filed: 01/11/2011
• Published: 05/12/2011
• Est. Priority Date: 12/16/2003
• Status: Active Grant

• Current Assignee: Precisense A/S (Medtronic Plc)
• Original Assignee: Precisense A/S (Medtronic Plc)
• Inventors: Laursen, Bo Wegge, Norrild, Jens Christian
• Granted Patent: US 8,318,443 B2
• Time in Patent Office: Days
• US Class Current: 436/172
• CPC Class Codes: C07C 309/14 containing amino groups bound to the carbon skeleton
• CPC Class Codes: C07D 207/404 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. succinimide
• CPC Class Codes: C07D 401/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C09B 11/10 Amino derivatives of triarylmethanes
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A reagent for use in detecting an analyte comprises a fluorescent energy donor and an energy acceptor, wherein the energy acceptor is of the general formula:

and wherein the distance between the energy donor and the energy acceptor of the reagent is capable of modulation by a suitable analyte to be detected.

Claims

1-32. -32. (canceled)

33. A reagent for use in detecting an analyte, comprising a fluorescent energy donor and an energy acceptor, the energy donor and the energy acceptor being such that when they are sufficiently close to one another energy is non-radiatively transferred from the energy donor following excitation thereof to the energy acceptor quenching fluorescence of the energy donor, wherein the energy acceptor is of the general formula:

34. A reagent as claimed in claim 33, wherein a linker structure is present, and is formed by reaction of a linker element selected from an active ester, an isothiocyanate, an acid chloride, an aldehyde, an azide, an α

-halogenated ketone and an amine with a reaction partner.

35. A reagent as claimed in claim 34, wherein the reaction partner is selected from a polysaccharide, a polynucleotide and a protein.

36. A reagent as claimed in claim 34, wherein the linker element is an active ester, and is selected from succinimidyl and pentafluorophenyl active esters.

37. A reagent as claimed in claim 34, wherein the energy donor is Alexa Fluor-594™

.

38. A dye compound having the general formula:

39. A dye compound as claimed in claim 38, wherein at least one of R¹⁰, R¹¹, R¹², R¹³, R¹⁴ and R¹⁵ is alkyl.

40. A dye compound as claimed in claim 39, wherein one or more pairs of groups R⁴ and R¹⁰ and/or R⁵ and R¹¹ and/or R⁶ and R¹² and/or R⁷ and R¹³ and/or R⁸ and R¹⁴ and/or R⁹ and R¹⁵ is a bridging group consisting of alkylene, O-alkylene, S-alkylene or N-alkylene optionally substituted with one or more of SO₃, PO₃²⁻

, OH, O-alkyl, SH, S-alkyl, COOH, COO⁻

, ester, amide, halogen, SO-alkyl, SO₂NH₂, SO₂NH-alkyl, SO₂N-dialkyl, SO₃-alkyl, CN, secondary amine or tertiary amine.