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:
CP 1 and CP 2 are independently selected from a 2,7-linked fluorene and a substituted 2,7-linked fluorene comprising replacement of a ring carbon atom of the 2,7-linked fluorene with a substituted heteroatom;
CP 3 is 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>0;
wherein the multichromophore is coupled to a signaling chromophore.
2. The multichromophore according to claim 1 , wherein the aromatic repeat unit is a carbocyclic or heterocyclic aryl group comprising 1 to 5 fused aromatic rings.
3. The multichromophore according to claim 2 , wherein the aromatic repeat unit is 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 2 , wherein the aromatic repeat unit is a heterocyclic aryl group having at least one heteroatom in at least one aromatic ring of an aryl group that is a 2,7-linked fluorene.
5. The multichromophore according to claim 2 , wherein:
the carbocyclic aryl group is a 2,7-linked fluorene; and
the heterocyclic aryl group has at least one heteroatom in at least one aromatic ring of an aryl group that is a 2,7-linked fluorene.
6. The multichromophore according to claim 1 , wherein the aromatic repeat unit is selected from benzene, naphthalene, anthracene, fluorene, thiophene, furan, pyridine, oxadiazole and substituted versions thereof.
7. The multichromophore according to claim 1 , wherein the multichromophore is substituted with a plurality of charge neutral water-soluble groups.
8. The multichromophore according to claim 1 , wherein CP 3 is present and comprises a bioconjugation site.
9. The multichromophore according to claim 1 , wherein CP 1 and CP 2 are substituted with one or more solubilizing groups selected from ethylene glycol oligomers, ethylene glycol polymers, ω-ammonium alkoxy salts and ω-sulfonate alkoxy salts.
10. The multichromophore according to claim 9 , wherein CP 1 , CP 2 and CP 3 are each substituted with —R 3 —C and —R 4 -D wherein:
R 3 and R 4 are independently selected from C1-22 alkyl, C1-22 alkoxy and polyalkylene oxide having from 1 to about 22 carbon atoms and C and D are each H.
11. The multichromophore according to claim 9 , wherein CP 1 , CP 2 and CP 3 are each substituted with one or more groups selected from —R 3 —C and —R 4 -D that are attached through a bridging functional group wherein:
R 3 and R 4 are independently selected from C1-22 alkyl, C1-22 alkoxy and polyalkylene oxide having from 1 to about 22 carbon atoms and C and D are each H; and
the bridging functional group is selected from —O—, —S—, —C(O)—, —C(O)O—, —C(R)(R′)—, —N(R′)—, and —Si(R′)(R″)—, wherein R, R′ and R″ are selected from hydrogen, C1-12 alkyl and C1-12 alkoxy.
12. The multichromophore according to claim 11 , wherein the bridging functional group is —Si(R′)(R″)—.