Photoactive macromolecules and uses thereof

A method for detecting an analyte in a sample comprising:

• providing a sample that is suspected of containing the analyte;
• contacting the sample with a binding agent conjugated to a water soluble fluorescent polymer having the structure of Formula I:
• wherein
• each X is C;
• each Y is CR1R2;
• each R1 is independently selected from the group consisting of PEG, ammonium alkyl salt, ammonium alkyloxy salt, ammonium oligoether salt, sulfonate alkyl salt, sulfonate alkoxy salt, sulfonate oligoether salt, sulfonamido oligoether, and
• each R2 is independently selected from the group consisting of H, alkyl, alkene, alkyne, cycloalkyl, haloalkyl, alkoxy, (hetero) aryloxy, aryl, (hetero) arylamino, a PEG group, ammonium alkyl salt, ammonium alkyloxy salt, ammonium oligoether salt, sulfonate alkyl salt, sulfonate alkoxy salt, sulfonate oligoether salt, sulfonamido oligoether, and
• each R3 is independently selected from the group consisting of H, alkyl, alkene, alkyne,
• cycloalkyl, haloalkyl, alkoxy, (hetero) aryloxy, aryl, (hetero) arylamino, and a PEG group;
• each Z is independently selected from the group consisting of CH2, O, and NH;
• each Q is independently selected from the group consisting of a bond, NH, NR4, and CH2;
• each M is independently an electron rich linker unit capable of altering the polymer band gap and is evenly or randomly distributed along the polymer main chain and is each independently selected from the group consisting of
• wherein
• each R4 is a non-ionic side group capable of imparting solubility in water in excess if of 10 mg/mL and is each independently selected from the group consisting of halogen, hydroxyl, C1-C12 alkyl, C2-C12 alkene, C2-C12 alkyne, C3-C12 cycloalkyl, C1-C12 haloalkyl, C1-C12 alkoxy, C2-C18 (hetero) aryloxy, C2-C18 (hetero) arylamino, (CH2)x′(OCH2—CH2)y′OCH3 where each x′ is independently an integer from 0-20;
• each y′ is independently an integer from 0-50, and a C2-C18 (hetero) aryl group;
• each optional linker L is independently an aryl or heteroaryl group evenly or randomly distributed along the polymer main chain and are substituted with one or more pendant chains terminated with a functional group selected from the group consisting of amine, carbamate, carboxylic acid, carboxylate, maleimide, activated ester, N-hydroxysuccinimidyl, hydrazine, hydrazid, hydrazone, azide, alkyne, aldehyde, thiol, and protected groups thereof for conjugation to another substrate, acceptor dye, molecule or binding agent, or conjugated to an acceptor dye or binding agent;
• G1 and G2 are each independently selected from the group consisting of hydrogen, halogen, alkyne, optionally substituted aryl, optionally substituted heteroaryl, halogen substituted aryl, silyl, diazonium salt, triflate, acetyloxy, azide, sulfonate, phosphate, boronic acid substituted aryl, boronic ester substituted aryl, boronic ester, boronic acid, optionally substituted dihydrophenanthrene (DHP), optionally substituted fluorene, aryl or heteroaryl substituted with one or more pendant chains terminated with a functional group selected from amine, carbamate, carboxylic acid, carboxylate, maleimide, activated ester, N-hydroxysuccinimidyl, hydrazine, hydrazide, hydrazone, azide, alkyne, aldehyde, thiol, and protected groups thereof for conjugation to a substrate or binding agent, or conjugated to a binding agent;
• a, c, and d define the mol % of each unit within the structure which each can be evenly or randomly repeated and where a is a mol % from 10 to 100%, c is a mol % from 0 to 90%, and each d is a mol % from 0 to 25%;
• each b is independently 0 or 1;
• m is an integer from 1 to about 10,000;
• and
• each n is independently an integer from 1 to 20;
• and
• wherein the binding agent is capable of interacting with the analyte or a target-associated biomolecule.