Peptide nucleic acid functionalized hydrogel microneedles for sampling and detection of interstitial fluid nucleic acids

Abstract

The present disclosure relates to a device, comprising a base and a plurality of microneedles attached to the base, wherein each microneedle has an outer surface; the outer surface of at least one microneedle being coated with a composition comprising at least one polymer and least one Peptide Nucleic Acid (PNA). The present disclosure additionally relates to a method of detecting an analyte in interstitial fluid (ISF), comprising contacting the device to a subject, for example, to human skin.

1 . A device for detecting an analyte, comprising a base, and a plurality of microneedles attached to the base, wherein:
 each microneedle has an outer surface; and   the outer surface of at least one microneedle is coated with a composition comprising at least one polymer and at least one first Peptide Nucleic Acid (PNA).   
     
     
         2 . The device of  claim 1 , wherein the composition further comprises at least one second PNA, wherein the second PNA is different from the first PNA. 
     
     
         3 . The device of  claim 1 , wherein the polymer is hydrophilic. 
     
     
         4 . The device of  claim 1 , wherein the polymer is alginate, xanthan, dextran, hyaluronic acid, poly(vinylalcohol) (PVA), polymethacrylic acid (PMAA), polyacrylic acid (PAA), poly(N-vinylpyrrolidone) (PVP), poly(lactic-co-glycolic acid) (PLGA), poly(N-isopropylacrylamide), poly(ethylene glycol) (PEG), poly(propylene oxide) (PPO), poly(ethylene glycol) diacrylate/dimethacrylate (PEGDA/PEGDMA), or poly(ethylene glycol) acrylate/methacrylate (PEGA/PEGMA), or a combination thereof. 
     
     
         5 . (canceled) 
     
     
         6 . The device of  claim 1 , wherein the polymer is covalently attached to the first PNA, optionally by a first linker. 
     
     
         7 . The device of  claim 2 , wherein the polymer is covalently attached to the second PNA, optionally by a second linker. 
     
     
         8 . The device of  claim 6  or  7 , wherein the linker is selected from  —OC(═O)— ,  (O═)CO— ,  —NH—C(═O)— ,  —C(═O)NH— ,  —O— ,  —NH—C(═O)—NH— , or  —S— , wherein   indicates a point of attachment of the linker to the polymer, to the first PNA, or to the second PNA; or wherein the linker is represented by structural formula (I) or (Ia), 
       
         
           
           
               
               
           
         
       
       wherein   indicates a point of attachment of the linker to the polymer, to the first PNA, or to the second PNA. 
     
     
         9 . (canceled) 
     
     
         10 . The device of  claim 6 , wherein the linker is cleavable. 
     
     
         11 . The device of  claim 10 , wherein the linker is a photocleavable linker represented by structural formula (II) or (IIa), 
       
         
           
           
               
               
           
         
       
       wherein   indicates a point of attachment of the linker to the polymer, to the first PNA, or to the second PNA. 
     
     
         12 . The device of  claim 1 , wherein the first PNA comprises from 5 to 30 nucleobases. 
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . The device of  claim 2 , wherein:
 the analyte is a nucleic acid;   the first PNA is complementary to the 5′ end of the nucleic acid; and   the second PNA is complementary to the 3′ end of the nucleic acid.   
     
     
         16 . (canceled) 
     
     
         17 . The device of  claim 6 , wherein the first PNA is represented by a structural formula selected from 
       
         
           
           
               
               
           
         
       
       wherein   indicates a point of attachment of the first PNA to the linker. 
     
     
         18 . The device of  claim 6 , wherein:
 the first PNA comprises a first probe head;   the second PNA comprises a second probe head; and   the first probe head and the second probe head selectively bind each other, thereby producing a detectable signal.   
     
     
         19 . The device of  claim 18 , wherein the first probe head comprises a chemical moiety selected from the group consisting of 
       
         
           
           
               
               
           
         
       
       and 
       the second probe head comprises a chemical moiety selected from the group consisting of 
       
         
           
           
               
               
           
         
       
       wherein: 
          is a point of attachment of the chemical moiety to the first PNA or to the second PNA; 
       each R or R′ is independently selected from Halogen, —NO 2 , —OH, —NH 2 , —NH(C 1-6  alkyl), —N(C 1-6  alkyl) 2 , —NCS, C 1-6  alkyl, C 1-6  haloalkyl, C 6-10  aryl, 5- to 12-membered heteroaryl, —O(C 1-6  alkyl), —C(O)O(C 1-6  alkyl), —OC(O)(C 1-6  alkyl), —C(O)NH 2 , —C(O)NH(C 1-6  alkyl), —C(O)N(C 1-6  alkyl) 2 , —NHC(O)(C 1-6  alkyl), —N(C 1-6  alkyl)C(O)(C 1-6  alkyl), —SO 2 (C 1-6  alkyl), —SO 2 (C 6-10  aryl), and —SO 3   − X + ; 
       X +  is Li + , Na + , K + , or N(C 1-6  alkyl) 4   + ; 
       m is 0 to 2; and 
       n is 0 to 4. 
     
     
         20 . The device of  claim 18 , wherein:
 the first probe head comprises a thiol and the second probe head comprises a chemical moiety represented by the structural formula (IV) or structural formula (V):   
       
         
           
           
               
               
           
         
       
       or
 the first probe head comprises a chemical moiety represented by the following structural formula 
 
       
         
           
           
               
               
           
         
       
       and the second probe head comprises a chemical moiety represented by the following structural formula 
       
         
           
           
               
               
           
         
       
       wherein   indicates a point of attachment of the chemical moiety to the first PNA or to the second PNA. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . The device of  claim 1 , wherein the first PNA is a hairpin PNA comprising a third probe head; and
 further wherein the third probe head produces a detectable signal upon the hairpin PNA binding to the analyte.   
     
     
         24 . The device of  claim 23 , wherein the third probe head comprises a chemical moiety selected from the group consisting of 
       
         
           
           
               
               
           
         
       
       wherein: 
          is a point of attachment of the chemical moiety to the first PNA or to the second PNA; 
       each R or R′ is independently selected from Halogen, —NO 2 , —OH, —NH 2 , —NH(C 1-6  alkyl), —N(C 1-6  alkyl) 2 , —NCS, C 1-6  alkyl, C 1-6  haloalkyl, C 6-10  aryl, 5- to 12-membered heteroaryl, —O(C 1-6  alkyl), —C(O)O(C 1-6  alkyl), —OC(O)(C 1-6  alkyl), —C(O)NH 2 , —C(O)NH(C 1-6  alkyl), —C(O)N(C 1-6  alkyl) 2 , —NHC(O)(C 1-6  alkyl), —N(C 1-6  alkyl)C(O)(C 1-6  alkyl), —SO 2 (C 1-6  alkyl), —SO 2 (C 6-10  aryl), and —SO 3   − X + ; 
       X +  is Li + , Na + , K + , or N(C 1-6  alkyl) 4   + ; and 
       n is 0 to 4. 
     
     
         25 . The device of  claim 18 , wherein the detectable signal is fluorescent signal or electrochemical signal. 
     
     
         26 . (canceled) 
     
     
         27 . A method of detecting an analyte in interstitial fluid (ISF) of a subject, comprising: contacting the subject with the device of  claim 18 ;
 exposing the device to the ISF of the subject;   detaching the device from the subject; and   measuring an intensity of the detectable signal.   
     
     
         28 . A method of detecting an analyte in ISF of a subject, comprising:
 contacting the subject with the device of  claim 1 , and   exposing the device to the ISF of the subject.   
     
     
         29 - 57 . (canceled)