Methods and processes for attaching compounds to matrices

Abstract

The present invention describes extremely rapid and efficient methods for the attachment of chemical moieties to matrices by the use of microwave technology. The methods of the invention can be applied in a variety of ways for the preparation of different types of matrices for a variety of applications including but not limited to the functionalization of various solid supports, and matrices in the form of powder, beads, sheets, and other suitable surfaces for use in applications including but not limited to oligonucleotide synthesis, peptide synthesis, environmental clean up (removal of toxic materials), immunoassays, affinity chromatography, combinatorial chemistry, microarrays, proteomics and medical diagnostics.

What is claimed is: 
     
         1 . A method for attaching a chemical moiety to a matrix comprising the steps of:
 (a) contacting the matrix with a reagent capable of adding a nucleophilic group;   (b) exposing the reaction mixture of step (a) to microwave radiation thereby resulting in a functionalized matrix;   (c) contacting the functionalized matrix of step (b) with a reagent capable of forming an ester or amide bond with the matrix and further comprising a free carboxyl termini on the matrix;   (d) exposing the reaction mixture of step (c) to microwave radiation thereby forming a mono-ester or mono-amide linkage with the matrix comprising a free carboxyl termini on the matrix; and   (e) coupling the carboxylated matrix of step (d) with the chemical moiety via a reative region of the chemical moiety capable of reacting with the carboxylated matrix thereby resulting in a matrix functionalized with the chemical moiety.   
     
     
         2 . The method of  claim 1  wherein the contacting of steps (a) and (c) are carried out in the presence of a solvent having a dielectric constant. 
     
     
         3 . The method of  claim 1  wherein the matrix is selected from the group consisting of: controlled pore glass; glass beads; glass powders; silica gels; alumina;
 substituted or unsubstituted polystyrene; polyethylene glycol; cellulose, ceramics, zeolite, clay, titanium (Ti), Carbon, silicon (Si), and gold. 
 
     
     
         4 . The method of  claim 1  wherein the chemical moiety is selected from the group consisting of: modified and unmodified nucleotides and nucleosides; DNA; RNA; amino acids; peptides; proteins; synthetic block polymers; small molecules; and organometallic synthesis reagents. 
     
     
         5 . A method for preparing a functionalized matrix for oligonucleotide synthesis comprising the steps of:
 (a) contacting the matrix with a reagent capable of adding an amino functional group to the matrix;   (b) exposing the reaction mixture of step (a) to microwave radiation thereby resulting in an amino-functionalized matrix;   (c) contacting the amino-functionalized matrix of step (b) with a succinylating reagent capable of chemically succinylating the matrix;   (d) exposing the reaction mixture of step (c) to microwave radiation thereby resulting in a succinylated matrix; and   (e) coupling the succinylated matrix with a nucleoside capable of reacting with the succinylated matrix thereby forming a functionalized matrix suitable for further use in the synthesis of oligonucleotides.   
     
     
         6 . The method of  claim 5  wherein the contacting of step (a) is carried out in the presence of a solvent having a dielectric constant. 
     
     
         7 . The method of  claim 5  wherein the reagent of step (a) capable of adding an amino functional group is an aminoalkylsilane. 
     
     
         8 . The method of  claim 6  wherein the solvent is dimethylformamide, dimethyl acetamide, N,N-dialkyl formamides and acetamides, N-methylpyrrolidone, and DMSO. 
     
     
         9 . The method of  claim 5  wherein the succinylating reagent is a substituted or unsubstituted dicarboxylic acid or their corresponding anhydrides, or any reagent capable of forming a mono-ester linkage with the matrix and having a free carboxyl termini. 
     
     
         10 . The method of  claim 9  wherein the succinylating reagent is succinic anhydride. 
     
     
         11 . The method of  claim 5  further comprising the step of recovering excess nucleoside generated in the coupling step (e) by aqueous work up of the filtrate. 
     
     
         12 . The method of  claim 5  wherein the matrix is selected from the group consisting of: controlled pore glass; glass beads; glass powders; silica gels; alumina;
 substituted or unsubstituted polystyrene; polyethylene glycol; cellulose, ceramics, zeolite, clay, titanium (Ti), Carbon, silicon (Si), and gold. 
 
     
     
         13 . The method of claim of  claim 5  wherein the matrix is controlled pore glass. 
     
     
         14 . The method of  claim 5  wherein the nucleoside derivative is a 5′-protected nucleoside derivative. 
     
     
         15 . The method of  claim 14  wherein the 5′ nucleoside derivative is 5′-dimethoxytrityl-protected nucleoside with a free 3′ hydroxyl group. 
     
     
         16 . The method of  claim 5  wherein the functionalized nucleoside matrix comprises a loading of nucleoside derivative in the range of about 60-100 micromoles of nucleoside derivative per gram of matrix. 
     
     
         17 . The method of  claim 5  wherein the amino-functionalized matrix of step (b) comprises a loading of amino group in the range of about 60-120 micromole of amino group per gram of matrix. 
     
     
         18 . The method of  claim 5  wherein the contacting of step (a) comprises contacting the matrix with at least two different reagents capable of adding amino functional groups to the matrix. 
     
     
         19 . A method of preparing a functionalized matrix for oligonucleotide synthesis comprising the steps of:
 (a) contacting the matrix with an aminoalkylsilane reagent in the presence of a solvent having a dielectric constant and exposing the reaction mixture to microwave radiation thereby resulting in an amino-functionalized matrix;   (b) reacting the matrix of step (a) with succinic anhydride in the presence of dimethylformamide and exposing the reaction mixture to microwave radiation thereby forming a succinylated matrix; and   (c) contacting the succinylated matrix of step (b) with a 5′ dimethoxytrityl-protected nucleoside derivative thereby forming a functionalized matrix suitable for further use in oligonucleotide synthesis.