UV excitable polyfluorene based conjugates and their use in methods of analyte detection

A method of detecting an analyte in a sample, comprising

• a) combining said sample with a detection reagent comprising a polymer under conditions under which said detection reagent will bind said analyte;
• and
• b) detecting the detection reagent bound analyte by fluorescence,
• wherein the polymer is optionally bound to an antibody,
• wherein the polymer comprises monomer units of formula A
• wherein X is the number of monomer units of formula A in the polymer wherein the monomer units of formula A are consecutive or nonconsecutive,
• and one or more monomer units of formula B
• wherein Y is the number of monomer units of formula B in the polymer wherein the monomer units of formula B are consecutive or nonconsecutive,
• and, optionally one or more monomer units of formula C
• wherein Z is the number of monomer units of formula C in the polymer wherein the monomer units of formula C are consecutive or nonconsecutive,
• wherein A is N;
• wherein SG1, SG2, SG5, SG6, each independently is a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein R1 is non-existent or a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein R2 is a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein SG3, SG4, R3 and R4 each independently is a hydrogen, a halogen, an amino, a PEG, a linker (L), an acceptor, or a biological substrate conjugated via a linker (L-BS);
• wherein the acceptor is a fluorophore or a fluorescent dye;
• wherein each polymer end independently is a hydrogen, an alkyl, a halogen, a boronyl, an aryl, a heteroaryl group or a L-BS;
• and
• wherein X, Y and Z are integers such that X+Y+Z is from 15 to 50, X is at least 10, and
• a ratio of X to Y+Z is 0.3-1.0.

A kit comprising

• a polymer comprising a conjugated biological substrate selected from the group consisting of a specific binding pair member, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid polymer, a peptide, and a protein to result in a labeled biological substrate, and
• one or more buffering agents,
• wherein the polymer comprises monomer units of formula A
• wherein X is the number of monomer units of formula A in the polymer wherein the monomer units of formula A are consecutive or nonconsecutive,
• and one or more monomer units of formula B
• wherein Y is the number of monomer units of formula B in the polymer wherein the monomer units of formula B are consecutive or nonconsecutive,
• and, optionally one or more monomer units of formula C
• wherein Z is the number of monomer units of formula C in the polymer wherein the monomer units of formula C are consecutive or nonconsecutive,
• wherein A is N;
• wherein SG1, SG2, SG5, SG6, each independently is a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein R1 is non-existent or a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein R2 is a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, an acceptor, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein SG3, SG4, R3 and R4 each independently is a hydrogen, a halogen, an amino, a PEG, a linker (L), an acceptor, or a biological substrate conjugated via a linker (L-BS);
• wherein the acceptor is a fluorophore or a fluorescent dye;
• wherein each polymer end independently is a hydrogen, an alkyl, a halogen, a boronyl, an aryl, a heteroaryl group or a L-BS;
• and
• wherein X, Y and Z are integers such that X+Y+Z is from 15 to 50, X is at least 10, and
• a ratio of X to Y+Z is 0.3-1.0.

A polymer comprising monomer units of formula A

• wherein X is the number of monomer units of formula A in the polymer wherein the monomer units of formula A are consecutive or nonconsecutive,
• and one or more monomer units of formula B
• wherein Y is the number of monomer units of formula B in the polymer wherein the monomer units of formula B are consecutive or nonconsecutive,
• and, optionally one or more monomer units of formula C
• wherein Z is the number of monomer units of formula C in the polymer wherein the monomer units of formula C are consecutive or nonconsecutive,
• wherein A is N;
• wherein SG1, SG2, SG5, SG6, each independently is a hydrogen, an alkyl, an amino, a sulfo, a polyethylene glycol (PEG), a water solubilizing group, a fluorophore or fluorescent dye, a linker (L), and/or a biological substrate conjugated via a linker (L-BS);
• wherein R1 is non-existent;
• wherein R2 is
• wherein SG3, SG4, R3 and R4 each independently is a hydrogen, a halogen, an amino, a PEG, a linker (L), an acceptor, or a biological substrate conjugated via a linker (L-BS);
• wherein the acceptor is a fluorophore or a fluorescent dye;
• wherein each polymer end independently is a hydrogen, an alkyl, a halogen, a boronyl, an aryl, a heteroaryl group or a L-BS;
• and
• wherein X, Y and Z are integers such that X+Y+Z is from 15 to 50, X is at least 10, and a ratio of X to Y+Z is 0.3-1.0.