Dyes having ratiometric fluorescence response for detecting metabolites
AssigneeBecton, Dickinson and Company
InventorsJoseph Thomas, Michael T. Cash
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Abstract
The presently disclosed subject matter provides thiol-reactive, environmentally sensitive fluorescent dyes, or fluorophores, which have an emission wavelength in the visible spectral region. When conjugated with a binding protein, the fluorophores exhibit a ratiometric response to one or more ligands or target analytes. The presently disclosed fluorophore-binding protein conjugates can be used to detect the presence of or amount of physiologically-important metabolites, such as glucose, fatty acids, and lactate, in biological samples.
Claims30
A fluorophore, wherein the fluorophore is selected from the group consisting of:
A biosensor compound having the following formula:
- wherein, B is a binding member having a binding affinity for a ligand or analyte to be detected
- wherein the biosensor compound exhibits a detectable change in a fluorescence property as a result of binding to the ligand or analyte or as a result of a change in concentration of the ligand or analyte in a sample under test.
A biosensor compound having the following formula:
- wherein:
- q′ is an integer from 1 to 8;
- each X1′ and X2′ is independently selected from the group consisting of C and N, wherein at least one of X1′ and X2′ is N, under the proviso that (i) when X1′ is C, R1′ is Z, or when X2′ is C, R2′ is Z, as Z is defined herein below;
- (ii) if both X1′ and X2′ are N at the same time, one of R1′ and R2′ is absent;
- and (iii) when X1′ is N, R1′ when present is Z″, or when X2′ is N, R2′ when present is Z″, wherein Z″ is
- —(CH2)n′X3′C(═O)—R4′,
- wherein:
- n′ is an integer from 1 to 8;
- X3′ is O or NR6′;
- each R3′, R6′, R7′, R8′, and Z is independently selected from the group consisting of H, alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, hydroxyalkyl, hydroxycycloalkyl, alkoxycycloalkyl, aminoalkyl, acyloxyl, alkylaminoalkyl, and alkoxycarbonyl;
- or
- R7′ and R8′ together represent a C2 to C10 alkyl, C2 to C10 substituted alkyl, or C2 to C10 alkylene;
- R4′ is —(CH2)m—B;
- wherein m′ is an integer from 1 to 8;
- and
- B is SM4, wherein SM4 is a mutated glucose/galactose binding protein (GGBP) having the following substitutions:
- N39I, G82E, Q83K, N84D, Q175E, Q177H, L178M, W183C, N259E and N260S.
Dependencies are inferred from the claim text.