Conjugated Polymeric Particle and Method of Making Same

Abstract

A method of conjugating a substrate includes exchanging a counter ion associated with a biomolecule with a lipophilic counter ion to form a biomolecule complex, dispersing the biomolecule complex in a nonaqueous solvent, and coupling the biomolecule complex to a substrate in the presence of the nonaqueous solvent.

1 .- 18 . (canceled) 
     
     
         19 . A method of isolating a target polynucleotide, the method comprising:
 contacting a first solution including the target polynucleotide with a probe conjugated substrate, the probe of the probe conjugated substrate being at least partially complementary to the target polynucleotide, the probe conjugated substrate formed by:
 exchanging a counter ion associated with an oligonucleotide with a lipophilic counter ion to form a biomolecule complex; 
 dispersing the biomolecule complex in a nonaqueous solvent; and 
 coupling the biomolecule complex to a polymeric particle in the presence of the nonaqueous solvent to form the oligonucleotide conjugated polymeric particle; 
   washing the probe conjugated substrate while the target polynucleotide is coupled to the probe; and   releasing the target polynucleotide in a second solution.   
     
     
         20 . The method of  claim 19 , wherein the lipophilic counter ion is a lipophilic ammonium ion, a lipophilic phosphonium ion, a lipophilic arsonium ion, a lipophilic sulfonium ion, or a combination thereof. 
     
     
         21 . The method of  claim 20 , wherein the lipophilic ammonium ion is a tetraalkylammonium, a tetraarylammonium, mixed alkyl and aryl ammonium, or a combination thereof. 
     
     
         22 . The method of  claim 21 , wherein the lipophilic ammonium ion is selected from the group consisting of tetramethylamonium, tetraethylamonium, tetrapropylamonium, tetrabutylamonium, tetrapentylamonium, tetrahexylamonium, tetraheptylamonium, tetraoctylamonium, alkyl and aryl mixtures thereof, and a combination thereof. 
     
     
         23 . The method of  claim 20 , wherein the lipophilic phosphonium ion is tetraphenylphosphonium. 
     
     
         24 . The method of  claim 20 , wherein the lipophilic arsonium ion is a tetraalkylarsonium, a tetraarylarsonium, a mixed alkyl and aryl arsonium ion, or a combination thereof. 
     
     
         25 . The method of  claim 24 , wherein the lipophilic arsonium ion is tetraphenylarsonium. 
     
     
         26 . The method of  claim 20 , wherein the lipophilic sulfonium ion is a trialkylsulfonium ion. 
     
     
         27 . The method of  claim 19 , wherein the nonaqueous solvent is non-reactive with coupling groups on the substrate and the biomolecule. 
     
     
         28 . The method of  claim 19 , wherein the nonaqueous solvent is polar. 
     
     
         29 . The method of  claim 19 , wherein the nonaqueous solvent is an amide, a urea, a carbonate, an ether, a sulfoxide, a sulfone, a hindered alcohol, or a combination thereof 
     
     
         30 . The method of  claim 29 , wherein the amide or urea is selected from a group consisting of formamide, N,N-dimethylformamide, acetamide, N,N-dimethylacetamide, hexamethylphosphoramide, pyrrolidone, N-methylpyrrolidone, N,N,N′,N′-tetramethylurea, N,N′-dimethyl-N,N′-trimethyleneurea, and a combination thereof. 
     
     
         31 . The method of  claim 29 , wherein the carbonate is selected from a group consisting of dimethyl carbonate, propylene carbonate, and a combination thereof. 
     
     
         32 . The method of  claim 29 , wherein the ether is tetrahydrofuran. 
     
     
         33 . The method of  claim 29 , wherein the sulfoxide or sulfone is selected from a group consisting of dimethylsulfoxide, dimethylsulfone, and a combination thereof. 
     
     
         34 . The method of  claim 29 , wherein the hindered alcohol includes tert-butyl alcohol. 
     
     
         35 . The method of  claim 19 , wherein the polymeric particle includes a coupling group reactive with a reactive group comprising the biomolecule. 
     
     
         36 . The method of  claim 35 , wherein the coupling group includes one of a nucleophile or an electrophile and the reactive partner includes the other of the nucleophile or the electrophile. 
     
     
         37 . The method of  claim 19 , wherein coupling the biomolecule complex to the polymeric particle includes:
 wherein the polymeric particle includes a polymer comprising amine functionality;   treating the polymer comprising amine functionality with a bis-NHS ester or a disuccinimidyl carbonate to form a functionalized polymer; and   treating the functionalized polymer with an amine terminated oligonucleotide to form a conjugated polymer including the oligonucleotide, the biomolecule complex including the amine terminated oligonucleotide.   
     
     
         38 . The method of  claim 37 , further comprising:
 wherein the polymeric particle includes an initial polymer including hydroxyl functionality;   treating the initial polymer to substitute a halogen for the hydroxyl functionality; and   further treating the initial polymer with a protected amine terminated polyether to form the polymer comprising the amine functionality.