Compositions and methods for separating heterocyclic aromatic amine bases, nucleosides, nucleotides, and nucleotide sequences

Abstract

The compositions of the present invention comprise one or more palladium bound ligands that are covalently bonded to inorganic or organic solid supports. These palladium bound ligands bonded to solid supports can be used for single heterocyclic amine base separation, or can be used to separate nucleotide chain containing specific sequences from other nucleotides or nucleotide chains. In one aspect of the invention, each ligand present is individually complexed to a single Pd(II) ion. If there are from 2 to 4 ligands present in the composition, then each ligand present must be separated from the other ligands by at least 3 atoms, preferably from 3 to 20 carbon atoms or equivalent spacing.

1 - 42 . (Canceled)  
     
     
         43 . A method for concentrating, removing, or separating a desired heterocyclic aromatic amine base or a nucleotide sequence containing said desired heterocyclic aromatic amine base from a source solution comprising the steps of: 
 (a) contacting a source solution having a first volume with a composition comprising a palladium(II)-bound ligand covalently bonded to a solid support, said source solution containing a desired heterocyclic aromatic amine base and at least one undesired heterocyclic aromatic amine base, wherein the palladium(II)-bound ligand is preferentially selective of the desired heterocyclic aromatic amine base;    (b) removing the source solution from contact with said composition to which said desired heterocyclic aromatic amine base has been complexed;    (c) contacting said composition having said desired heterocyclic aromatic amine bases associated therewith with a smaller volume of an aqueous receiving solution in which said desired heterocyclic aromatic amine bases are stripped from the composition; and    (d) recovering said desired heterocyclic aromatic amine bases in concentrated form in said receiving solution.    
     
     
         44 . A method as in  claim 43  wherein said desired heterocyclic aromatic amine base is guanine (G), and the ligand portion of the palladium(II)-bound ligand comprises the sequence  
         —[B—C x —B—(C x —B) y ] 
       wherein C is carbon, each B is independently nitrogen, sulfur, or phosphene; x is 2 or 3; and y is 0, 1 or 2.  
     
     
         45 . A method as in  claim 43  wherein said desired heterocyclic aromatic amine base is adenine (A), and the ligand portion of the palladium(II)-bound ligand comprises the sequence  
         -[D-C—N—C-D] 
       wherein C is carbon, N is nitrogen, and D is a negative binder.  
     
     
         46 . A method as in  claim 43 , wherein said desired heterocyclic aromatic amine base is thymine (T), and the ligand portion of the palladium(II)-bound ligand comprises the sequence  
       
         
           
           
               
               
           
         
       
     
     
         47 . A method as in  claim 43 , wherein said desired heterocyclic aromatic amine base is uracil (U), and the ligand portion of the palladium(II)-bound ligand comprises the sequence  
       
         
           
           
               
               
           
         
       
     
     
         48 . A method as in  claim 43 , wherein said desired heterocyclic aromatic amine base is cytosine (C), and the ligand portion of the palladium(II)-bound ligand comprises the sequence  
       
         
           
           
               
               
           
         
       
     
     
         49 . A method as in  claim 43 , wherein the aqueous receiving solution is desired from the group consisting of ammonia, ammonia buffer, ethylene diamine, and thiocyanate.  
     
     
         50 . A method as in  claim 49 , wherein the aqueous receiving solution is ammonia or ammonia buffer.  
     
     
         51 . A method as in  claim 43 , wherein the aqueous receiving solution is strong enough to displace the heterocyclic aromatic amine base from the composition without altering its configuration and is not strong enough to displace the Pd(II) from the composition.  
     
     
         52 . A method as in  claim 43 , further comprising a washing step to remove the aqueous receiving solution from the composition for further use.  
     
     
         53 . A method as in  claim 43 , wherein the composition is configured for separating individual heterocyclic aromatic amine bases.  
     
     
         54 . A method as in  claim 43 , wherein the composition is configured for separating heterocyclic aromatic amine bases that are part of nucleotide sequences.  
     
     
         55 . A method for concentrating, removing, or separating a nucleotide chain containing a specific nucleotide sequence from a source solution comprising the steps of: 
 (a) identifying a nucleotide chain that contains a desired nucleotide sequence that is desired for concentrating, removing, or separating from a source solution;    (b) providing a composition that will selectively bind the nucleotide sequence;    (c) contacting the source solution having a first volume with the composition wherein a chemical attraction between the composition and the nucleotide sequence occurs;    (d) removing the source solution from contact with the composition, said composition having the nucleotide sequence attracted thereto;    (e) contacting the composition having the nucleotide sequence attracted thereto with a smaller volume of an aqueous receiving solution in which said nucleotide chain is stripped from the composition; and    (f) recovering the nucleotide chain in concentrated form in the receiving solution.    
     
     
         56 . A method as in  claim 55 , wherein the nucleotide sequence contains from 2 to 4 heterocyclic amine bases.  
     
     
         57 . A method as in  claim 55 , wherein the source solution contains other non-desired nucleotide sequences are present in the source solution  
     
     
         58 . A method as in  claim 55 , wherein the composition has from 2 to 4 palladium(II)-bound ligands, said palladium(II)-bound ligands being tethered together by an organic backbone having from 3 to 20 atoms between each palladium(II)-bound ligand, thereby forming a palladium(II)-bound ligand backbone grouping, said palladium(II)-bound ligand backbone grouping covalently bonded to a solid support, wherein each palladium(II)-bound ligand is independently selective of a desired heterocyclic aromatic amine base of the nucleotide sequence.  
     
     
         59 . A method as in  claim 55 , wherein the nucleotide chain is the same as the nucleotide sequence.  
     
     
         60 . A method as in  claim 55 , wherein the nucleotide chain is longer than the nucleotide sequence.  
     
     
         61 - 67 . (Canceled)