Fluorescent methods and materials for directed biomarker signal amplification
Methods and compositions are provided that include a multichromophore and/or multichromophore complex for identifying a target biomolecule. A sensor biomolecule, for example, an antibody can be covalently linked to the multichromophore. Additionally, a signaling chromophore can be covalently linked to the multichromophore. The arrangement is such that the signaling chromophore is capable of receiving energy from the multichromophore upon excitation of the multichromophore. Since the sensor biomolecule is capable of interacting with the target biomolecule, the multichromophore and/or multichromophore complex can provide enhanced detection signals for a target biomolecule.
1. A water soluble multichromophore, comprising the structure:
wherein:
each CP 1 is selected from a carbocyclic aryl group that is a fluorene and a heterocyclic aryl group having at least one heteroatom in at least one aromatic ring of an aryl group that is a fluorene;
CP 2 and CP 3 are each independently an aromatic repeat unit;
m and n are independently 0 to 10,000 wherein m+n>1; and
a, b and c are independently 0 to 250 wherein a+b+c>1;
wherein CP 1 is present and each CP 1 and CP 2 are independently substituted with one or more water solubilizing groups selected from ethylene glycol oligomers and ethylene glycol polymers; and
CP 3 is present and is substituted with a linker to a bioconjugation functional group capable of covalently linking to a biomolecule or dye, wherein the bioconjugation functional group is selected from the group consisting of maleimide, thiol, succinimidyl ester, amine, azide and carboxy/EDC (1-Ethyl-3-[dimethylaminopropyl]carbodiimide hydrochloride).
2. The multichromophore according to claim 1 , wherein the aromatic repeat units are each independently a carbocyclic or heterocyclic aryl group comprising 1 to 5 fused aromatic rings.
3. The multichromophore according to claim 2 , wherein the aromatic repeat units are each independently a carbocyclic or heterocyclic aryl group, wherein the aryl group is selected from benzene, naphthalene, anthracene, fluorene, thiophene, furan, pyridine, oxadiazole and substituted versions thereof.
4. The multichromophore according to claim 1 , wherein the multichromophore is further coupled to a signaling chromophore.
5. The multichromophore according to claim 1 , wherein CP 1 and CP 2 are present and independently selected from a fluorene and a substituted fluorene comprising replacement of a carbon atom with a substituted heteroatom.
6. An aqueous composition comprising a water soluble multichromophore, comprising the structure:
wherein:
each CP 1 is selected from a carbocyclic aryl group that is a fluorene and a heterocyclic aryl group having at least one heteroatom in at least one aromatic ring of an aryl group that is a fluorene;
CP 2 and CP 3 are each independently an aromatic repeat unit;
m and n are independently 0 to 10,000 wherein m+n>1; and
a, b and c are independently 0 to 250 wherein a+b+c>1;
wherein CP 1 is present and each CP 1 and CP 2 are independently substituted with one or more water solubilizing groups selected from ethylene glycol oligomers and ethylene glycol polymers; and
CP 3 is present and is substituted with a linker to a bioconjugation functional group capable of covalently linking to a biomolecule or dye, wherein the bioconjugation functional group is selected from the group consisting of maleimide, thiol, succinimidyl ester, amine, azide and carboxy/EDC (1-ethyl-3-[dimethylaminopropyl]carbodiimide hydrochloride).
7. The aqueous composition according to claim 6 , wherein the aromatic repeat units are each independently a carbocyclic or heterocyclic aryl group, wherein the aryl group is selected from benzene, naphthalene, anthracene, fluorene, thiophene, furan, pyridine, oxadiazole and substituted versions thereof.
8. The aqueous composition according to claim 6 , wherein the multichromophore is further coupled to a signaling chromophore.
9. The aqueous composition according to claim 6 , wherein CP 1 and CP 2 are present and independently selected from a fluorene and a substituted fluorene comprising replacement of a carbon atom with a substituted heteroatom.