Methods and systems for analysis using polymer dots

Abstract

Methods, systems, compositions and kits are provided for the analysis of target molecules using chromophoric polymer dots conjugated to biomolecules. The use of chromophoric polymer dots improves detection sensitivity and stability when compared with existing techniques. In some aspects, methods, systems, and kits are provided for detecting a target protein using chromophoric polymer dots conjugated to biomolecules in a Western blot analysis. Related methods, systems, compositions and kits are also provided.

What is claimed is: 
     
         1 . A system for assaying a protein or peptide, the system comprising:
 a polymer dot conjugated to a biomolecule;   a protein or peptide separation platform;   a source of electromagnetic radiation;   a detector; and   a computer comprising a memory device with executable instructions stored thereon, the instructions, when executed by the processor, cause the processor to:
 operate the detector to acquire an image, and 
 store the image. 
   
     
     
         2 . The system of  claim 1 , wherein the polymer dot comprises a hydrophobic semiconducting polymer. 
     
     
         3 . The system of  claim 2 , wherein the hydrophobic semiconducting polymer is collapsed into a stable sub-micron-sized particle. 
     
     
         4 . The system of  claim 1 , further comprising a hydrophobic membrane. 
     
     
         5 . The system of  claim 4 , wherein the hydrophobic membrane comprises a nitrocellulose membrane or a fluoropolymer membrane. 
     
     
         6 . The system of  claim 1 , wherein the protein or peptide separation platform comprises a gel separation platform or a capillary electrophoresis separation platform. 
     
     
         7 . The system of  claim 1 , wherein the biomolecule is a protein, a nucleic acid, a peptide, or a lipid. 
     
     
         8 . The system of  claim 1 , further comprising a spectral filter, a multichroic mirror, or a combination thereof. 
     
     
         9 . The system of  claim 1 , wherein the detector comprises a gel imaging apparatus, a microscope, a camera, or a combination thereof. 
     
     
         10 . The system of  claim 1 , wherein the polymer dot comprises a BODIPY derivative having the structure of Formula 
       
         
           
           
               
               
           
         
         wherein each of R 1 , R 2A , R 2B , R 3A , R 4A  and R 4B  is independently selected from hydrogen, alkyl, aralkyl, aryl, and alkoxy-aryl, and wherein the BODIPY derivative is integrated into the chromophoric polymer by attachment to R 1 , R 2A , R 2B , R 3A , R 4A  and R 4B , or a combination thereof. 
       
     
     
         11 . The system of  claim 1 , wherein the semiconducting polymer comprises a narrow-band emissive unit. 
     
     
         12 . The system of  claim 11 , wherein the narrow-band emissive unit comprises a BODIPY monomer, a squaraine monomer, a metal complex, a porphyrin, a phthalocyanine, a lanthanide complex, a perylene, a cyanine, a rhodamine, a couramin, a xanthene, or any derivative thereof. 
     
     
         13 . The system of  claim 11 , wherein the narrow-band emissive unit is covalently attached to the semiconducting polymer. 
     
     
         14 . The system of  claim 1 , wherein the polymer dot comprises a functional group suitable for bioconjugation attached to the polymer dot. 
     
     
         15 . A kit for performing Western blot analysis, the kit comprising polymer dots, a conjugation biomolecule, and a blocking agent. 
     
     
         16 . The kit of  claim 15 , wherein the polymer dots comprise a hydrophobic semiconducting polymer collapsed into a stable sub-micron-sized particle. 
     
     
         17 . The kit of  claim 15 , wherein the polymer dots comprise a BODIPY derivative having the structure of Formula 
       
         
           
           
               
               
           
         
         wherein each of R 1 , R 2A , R 2B , R 3A , R 4A  and R 4B  is independently selected from hydrogen, alkyl, aralkyl, aryl, and alkoxy-aryl, and wherein the BODIPY derivative is integrated into the chromophoric polymer by attachment to R 1 , R 2A , R 2B , R 3A , R 4A  and R 4B , or a combination thereof. 
       
     
     
         18 . The kit of  claim 15 , wherein the polymer dots comprise a functional group attached to the polymer dot suitable for bioconjugation, 
     
     
         19 . The kit of  claim 15 , further comprising a hydrophobic membrane. 
     
     
         20 . The kit of  claim 15 , wherein the hydrophobic membrane is selected from a nitrocellulose membrane or a fluoropolymer membrane.