Methods and devices for removal of organic molecules from biological mixtures using anion exchange

Abstract

Methods and devices for removing small negatively charged molecules from a biological sample mixture that uses an anion exchange material that has associated therewith a polyoxyalkylene.

1 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a surface comprising an anion exchange material having associated therewith a polyoxyalkylene;    providing a biological sample mixture comprising small negatively charged organic molecules having a molecular weight of less than about 6,000; wherein the biological sample mixture is selected from the group consisting of a nucleic acid amplification reaction mixture and a nucleic acid labeling reaction mixture; and    contacting the biological sample mixture with the surface comprising the anion exchange material to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture.    
     
     
         2 . The method of  claim 1  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         3 . The method of  claim 2  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         4 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a surface comprising an anion exchange material partially coated with a negatively charged polymer, wherein the anion exchange material has associated therewith a polyoxyalkylene;    providing a biological sample mixture; and    contacting the biological sample mixture with the surface comprising an anion exchange material partially coated with a negatively charged polymer to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture.    
     
     
         5 . The method of  claim 4  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         6 . The method of  claim 5  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         7 . The method of  claim 4  wherein the negatively charged polymer is a polyelectrolyte.  
     
     
         8 . The method of  claim 7  wherein the negatively charged polyelectrolyte is selected from the group consisting of a polystryene sulfonic acid, polyvinyl phosphonic acid, polyvinyl boric acid, polyvinyl sulfonic acid, polyvinyl sulfuric acid, polystyrene phosphonic acid, polyacrylic acid, polymethacrylic acid, lignosulfonate, carrageenan, heparin, chondritin sulfate, salts thereof, and mixtures thereof.  
     
     
         9 . The method of  claim 4  wherein the anion exchange material comprises quaternized nitrogen.  
     
     
         10 . The method of  claim 4  wherein the biological sample mixture is a nucleic acid sequencing reaction mixture.  
     
     
         11 . The method of  claim 10  wherein the small negatively charged organic molecules are selected from the group consisting of dye-labeled terminators, primers, degraded dye molecules, deoxynucleotide triphosphates, and mixtures thereof.  
     
     
         12 . The method of  claim 11  wherein the small negatively charged organic molecules comprise dye-labeled terminators.  
     
     
         13 . The method of  claim 12  wherein the dye-labeled terminators are selected from the group consisting of dideoxynucleotide triphosphates, dideoxynucleotide diphosphates, dideoxynucleotide monophosphates, dideoxynucleosides, and combinations thereof.  
     
     
         14 . The method of  claim 13  wherein contacting the biological sample mixture with the surface comprising an anion exchange material partially coated with a negatively charged polymer is carried out under conditions effective to remove substantially all the dye-labeled terminators from the biological sample mixture.  
     
     
         15 . The method of  claim 4  wherein the biological sample mixture is a PCR reaction mixture.  
     
     
         16 . The method of  claim 15  wherein the small negatively charged organic molecules are selected from the group consisting of primers, degraded dye molecules, deoxynucleotide triphosphates, and mixtures thereof.  
     
     
         17 . The method of  claim 16  wherein contacting the biological sample mixture with the surface comprising an anion exchange material partially coated with a negatively charged polymer is carried out under conditions effective to remove substantially all the primers from the biological sample mixture.  
     
     
         18 . The method of  claim 4  wherein the small negatively charged organic molecules have a molecular weight of less than about 6,000.  
     
     
         19 . The method of  claim 4  wherein contacting the biological sample mixture with the surface comprising an anion exchange material partially coated with a negatively charged polymer comprises agitating while contacting.  
     
     
         20 . The method of  claim 4  which is carried out in a microfluidic device.  
     
     
         21 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a surface comprising an anion exchange material comprising quaternary ammonium ions partially coated with a negatively charged polyelectrolyte, wherein the anion exchange material has associated therewith a polyoxyalkylene;    providing a biological sample mixture; and    contacting the biological sample mixture with the surface comprising quaternary ammonium ions partially coated with a negatively charged polyelectrolyte to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture.    
     
     
         22 . The method of  claim 21  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         23 . The method of  claim 22  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         24 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a surface comprising an anion exchange material comprising quaternary ammonium ions partially coated with a negatively charged polyelectrolyte, wherein the anion exchange material has associated therewith a polyoxyalkylene;    providing a biological sample mixture; and    contacting the biological sample mixture with the surface comprising quaternary ammonium ions partially coated with a negatively charged polyelectrolyte to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture;    wherein the biological sample mixture comprises a nucleic acid amplification reaction mixture.    
     
     
         25 . The method of  claim 24  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         26 . The method of  claim 25  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         27 . The method of  claim 24  which is carried out in a microfluidic device.  
     
     
         28 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a device comprising at least one process array that comprises a surface comprising an anion exchange material having associated therewith a polyoxyalkylene;    providing a biological sample mixture in the at least one process array, wherein the biological sample mixture comprises small negatively charged organic molecules having a molecular weight of less than about 6,000; and    transferring the biological sample mixture within the at least one process array, wherein the biological sample mixture and the surface comprising an anion exchange material remain in contact for a sufficient time to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture.    
     
     
         29 . The method of  claim 28  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         30 . The method of  claim 29  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         31 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a device comprising at least one process array that comprises a surface comprising an anion exchange material partially coated with a negatively charged polymer, wherein the anion exchange material has associated therewith a polyoxyalkylene;    providing a biological sample mixture in the at least one process array; and    transferring the biological sample mixture within the at least one process array, wherein the biological sample mixture and the surface comprising an anion exchange material partially coated with a negatively charged polymer remain in contact for a sufficient time to remove at least a portion of the small negatively charged organic molecules from the biological sample mixture.    
     
     
         32 . The method of  claim 31  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         33 . The method of  claim 32  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         34 . The method of  claim 31  wherein the negatively charged polymer is a polyelectrolyte.  
     
     
         35 . The method of  claim 34  wherein the negatively charged polyelectrolyte is selected from the group consisting of a polystryene sulfonic acid, polyvinyl phosphonic acid, polyvinyl boric acid, polyvinyl sulfonic acid, polyvinyl sulfuric acid, polystyrene phosphonic acid, polyacrylic acid, polymethacrylic acid, lignosulfonate, carrageenan, heparin, chondritin sulfate, salts thereof, and mixtures thereof.  
     
     
         36 . The method of  claim 31  wherein the anion exchange material comprises quaternary ammonium ions.  
     
     
         37 . The method of  claim 31  wherein the biological sample mixture is a nucleic acid sequencing reaction mixture.  
     
     
         38 . The method of  claim 37  wherein the small negatively charged organic molecules are selected from the group consisting of dye-labeled terminators, primers, degraded dye molecules, deoxynucleotide triphosphates, and mixtures thereof.  
     
     
         39 . The method of  claim 38  wherein the small negatively charged organic molecules comprise dye-labeled terminators.  
     
     
         40 . The method of  claim 39  wherein the dye-labeled terminators are selected from the group consisting of dideoxynucleotide triphosphates, dideoxynucleotide diphosphates, dideoxynucleotide monophosphates, dideoxynucleosides, and combinations thereof.  
     
     
         41 . The method of  claim 40  wherein the biological sample mixture and the surface comprising an anion exchange material partially coated with a negatively charged polymer are contacted under conditions effective to remove substantially all the dye-labeled terminators from the biological sample mixture.  
     
     
         42 . The method of  claim 31  wherein the biological sample mixture is a PCR reaction mixture.  
     
     
         43 . The method of  claim 42  wherein the small negatively charged organic molecules are selected from the group consisting of primers, degraded dye molecules, deoxynucleotide triphosphates, and mixtures thereof.  
     
     
         44 . The method of  claim 43  wherein the biological sample mixture and the surface comprising an anion exchange material partially coated with a negatively charged polymer are contacted under conditions effective to remove substantially all the primers from the biological sample mixture.  
     
     
         45 . The method of  claim 31  wherein the small negatively charged organic molecules have a molecular weight of less than about 6,000.  
     
     
         46 . The method of  claim 31  wherein the biological sample mixture and the surface comprising an anion exchange material partially coated with a negatively charged polymer are agitated while in contact.  
     
     
         47 . The method of  claim 31  wherein the at least one process array comprises a loading chamber, at least one process chamber, and at least one distribution channel connecting the loading chamber and the at least one process chamber.  
     
     
         48 . A method of removing small negatively charged organic molecules from a biological sample mixture, the method comprising: 
 providing a device comprising at least one process array that comprises a surface comprising an anion exchange material comprising quaternary ammonium ions partially    
     
     
         54 . The device of claim  53  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         55 . The device of  claim 54  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         56 . The device of claim  53  further comprising a plurality of valves, wherein at least one of the valves is located along the at least one distribution channel.  
     
     
         57 . The device of claim  53  wherein the plurality of process arrays comprises a plurality of independent process arrays.  
     
     
         58 . The device of claim  53  wherein the plurality of process arrays are arranged radially on the device.  
     
     
         59 . The device of claim  53  wherein the surface comprising an anion exchange material comprises an anion exchange material partially coated with a negatively charged polymer.  
     
     
         60 . The device of  claim 59  wherein the negatively charged polymer is a polyelectrolyte.  
     
     
         61 . The device of  claim 59  wherein the anion exchange material, the negatively charged polymer, or both are pattern coated.  
     
     
         62 . An analytical receptacle comprising one or more reservoirs and a surface with a cover film adhered to the surface and enclosing the one or more reservoirs; wherein the cover film comprises a backing and an adhesive disposed on at least one major surface of the backing and in contact with the receptacle surface; wherein at least a portion of the adhesive has an anion exchange material disposed thereon; wherein the anion exchange material has associated therewith a polyoxyalkylene.  
     
     
         63 . The analytical receptacle of  claim 62  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         64 . The analytical receptacle of  claim 63  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         65 . The analytical receptacle of  claim 62  wherein the anion exchange material comprises quaternary ammonium ions.  
     
     
         66 . The analytical receptacle of  claim 62  wherein the anion exchange material is partially coated with a negatively charged polymer.  
     
     
         67 . The analytical receptacle of  claim 66  wherein the negatively charged polymer is a polyelectrolyte.  
     
     
         68 . The analytical receptacle of  claim 66  wherein the anion exchange material, the negatively charged polymer, or both are pattern coated.  
     
     
         69 . An analytical receptacle comprising a plurality of reservoirs adapted for receipt of a biological sample mixture, wherein at least one reservoir comprises a surface comprising an anion exchange material partially coated with a negatively charged polymer disposed therein, wherein the anion exchange material has associated therewith a polyoxyalkylene.  
     
     
         70 . The analytical receptacle of  claim 69  wherein the polyoxyalkylene is a nucleophilic polyoxyalkylene.  
     
     
         71 . The analytical receptacle of  claim 70  wherein the nucleophilic polyoxyalkylene is crosslinked.  
     
     
         72 . The analytical receptacle of  claim 69  wherein the anion exchange material comprises quaternary ammonium ions and the negatively charged polymer is a negatively charged polyelectrolyte.  
     
     
         73 . The analytical receptacle of  claim 69  which is a multi-well plate.  
     
     
         74 . A substrate comprising a functionalized surface on which is disposed an anion exchange material partially coated with a negatively charged polymer, wherein the anion exchange material has associated therewith a polyoxyalkylene.  
     
     
         75 . The substrate of  claim 74  wherein the anion exchange material and the polyoxyalkylene are mixed together.  
     
     
         76 . The substrate of  claim 74  wherein the anion exchange material is overcoated with the polyoxyalkylene, which is partially overcoated with the negatively charged polymer.  
     
     
         77 . The substrate of  claim 74  wherein the polyoxyalkylene is overcoated with the anion exchange material, which is partially overcoated with the negatively charged polymer.