Nanoparticle polyanion conjugates and methods of use thereof in detecting analytes

Abstract

This invention provides polyanionic polymer conjugates containing non-nucleotide polyanionic polymers that are useful in detecting target analytes such as proteins or small molecules. The invention also provides nanoparticles bound to polyanionic polymer conjugates and methods of preparation and use thereof. The polyanionic polymer conjugates have the formula: L-O—[PO 2 —O-Z-O] n —PO 2 —O—X wherein n ranges from 1 to 200; L represents a moiety comprising a functional group for attaching the polyanion polymer to the nanoparticle surface; Z represents a bridging group, and X represents Q, X′ or -Q-X′, wherein Q represents a functional group for attaching a recognition probe to the polyanion polymer, and X′ represents a recognition probe.

1 - 9 . (canceled) 
     
     
         10 . A nanoparticle having a plurality of polyanionic polymer conjugates attached thereto, said polyanionic polymer conjugates having the formula L-O—[PO 2 —O-Z-O] n —PO 2 —O—X wherein n ranges from 1 to 200; L represents a moiety comprising a functional group for attaching the polyanion polymer to a nanoparticle surface wherein the polyanion polymer is a non-nucleotide Polyanion polymer; Z represents a bridging group, and X represents Q, X′ or -Q-X′, wherein Q represents a functional group for attaching a recognition probe to the polyanion polymer, and X′ represents a recognition probe. 
     
     
         11 . The nanoparticle of  claim 10 , wherein the polyanionic polymer conjugate further comprises a detection label bound thereto. 
     
     
         12 . The nanoparticle of  claim 11 , wherein the detection label comprises a chromophore, a fluorescent label, a UV label, a radioisotope, a Raman label or a SERS (surface enhanced raman spectroscopy) label, or an enzyme. 
     
     
         13 . The nanoparticle of  claim 10 , wherein the functional group for attaching a recognition probe to the polyanion polymer comprises a carboxylic acid or an amino group. 
     
     
         14 . The nanoparticle of  claim 10 , wherein the recognition probe comprises a protein, a peptide, a nucleic acid, a peptide nucleic acid, a linked nucleic acid, a nucleoside triphosphate, a carbohydrate, a lipid, a lipid bound protein, an aptamer, a virus, a cell fragment, or a whole cell. 
     
     
         15 . The nanoparticle of  claim 14 , wherein the lipid bound protein comprises a G-protein coupled receptor. 
     
     
         16 . The nanoparticle of  claim 10 , wherein the recognition probe comprises an antibody, an antigen, a receptor, or a ligand. 
     
     
         17 . The nanoparticle of  claim 10  wherein L comprises an alkanethiol containing group, a phosphorothioate containing group, a substituted alkylsiloxane containing group, a polythiol containing group, or a cyclic disulfide containing group. 
     
     
         18 . The nanoparticle of  claim 10  wherein Z comprises a polymer, —C 1 -C 10 -alkyl-, —COO—, —CH 2 (CH 2 ) v COO—, —OCO—, R 1 N(CH 2 ) v —NR 1 —, —OC(CH 2 ) v —, —(CH 2 ) v —, —O—(CH 2 ) v —O—, —R 1 N—(CH 2 ) v —, 
       
         
           
           
               
               
           
         
         or 
       
       
         
           
           
               
               
           
         
         v is 0-30 and R 1  is H or is G(CH 2 ) v , wherein G is —CH 3 , —CHCH 3 , —COOH, —CO 2 (CH 2 ) v CH 3 , —OH, or —CH 2 OH. 
       
     
     
         19 - 42 . (canceled) 
     
     
         43 . A kit for detecting the presence or absence of a target analyte in a sample comprising:
 (a) nanoparticles having polyanionic polymer conjugates bound thereto, wherein the polyanion polymers have the formula:
   L-O—[PO 2 —O-Z-O] n —PO 2 —O—X 
   
       wherein n ranges from 1 to 200; L represents a moiety comprising a functional group for attaching the polyanion polymer to a nanoparticle surface wherein the polyanion polymer is a non-nucleotide polyanion polymer; Z represents a bridging group, and X represents Q, X′ or -Q-X′, wherein Q represents a functional group for attaching a recognition probe to the polyanion polymer, and X′ represents a recognition probe; and
 (b) an optional substrate for observing a detectable change. 
 
     
     
         44 . The kit of  claim 43 , wherein the polyanionic polymer conjugate further comprises a detection label bound thereto. 
     
     
         45 . The kit of  claim 44 , wherein the detection label comprises a chromophore, a fluorescent label, a UV label, a radioisotope, a Raman label or a SERS (surface enhanced raman spectroscopy) label, or an enzyme. 
     
     
         46 . The kit of  claim 43  wherein the functional group for attaching a recognition probe to the polyanion polymer comprises a carboxylic acid or an amino group. 
     
     
         47 . The kit of  claim 43 , wherein the recognition probe comprises a protein, a peptide, a nucleic acid, a peptide nucleic acid, a linked nucleic acid, a nucleoside triphosphate, a carbohydrate, a lipid, a lipid bound protein, an aptamer, a virus, a cell fragment, or a whole cell. 
     
     
         48 . The kit of  claim 47 , wherein the lipid bound protein comprises a G-protein coupled receptor. 
     
     
         49 . The kit of  claim 43 , wherein the recognition probe comprises an antibody, an antigen, a receptor, or a ligand. 
     
     
         50 . The kit of  claim 43  wherein the substrate is a transparent substrate or an opaque white substrate. 
     
     
         51 . The kit of  claim 43  wherein L comprises an alkanethiol containing group, a phosphorothioate containing group, a substituted alkylsiloxane containing group, a polythiol containing group, or a cyclic disulfide containing group. 
     
     
         52 . The kit of  claim 43  wherein Z comprises a polymer, —C 1 -C 10 -alkyl-, —COO—, —CH 2 (CH 2 ) v COO—, —OCO—, R 1 N(CH 2 ) v —NR 1 —, —OC(CH 2 ) v —, —(CH 2 ) v —, —O—(CH 2 ) v —O—, —R 1 N—(CH 2 ) v —, 
       
         
           
           
               
               
           
         
         or 
       
       
         
           
           
               
               
           
         
         v is 0-30 and R 1  is H or is G(CH 2 ) v , wherein G is —CH 3 , —CHCH 3 , —COOH, —CO 2 (CH 2 ) v CH 3 , —OH, or —CH 2 OH.