Nonaqueous polymeric reagent compositions and applications thereof

• US 5,260,195 A
• Filed: 01/03/1991
• Issued: 11/09/1993
• Est. Priority Date: 01/03/1991
• Status: Expired due to Fees

• Current Assignee: Roche Diabetes Care Inc. (Roche Holding AG)
• Original Assignee: Boehringer Mannheim Corporation
• Inventors: Rapkin, Myron C., DuBois-Bousamra, Jill, Skarstedt, Mark T., Burke, Anthony D., Azhar, Abol F., Diebold, Eric R., Usmani, Arthur M.
• Primary Examiner(s): Griffin, Ronald W.
• Assistant Examiner(s): Webber, Pamela S.
• Application Number:
  US07/638,263
• Time in Patent Office: 1,041 Days
• Field of Search: 435/188, 435/805, 435/25, 435/28, 436/166, 436/518, 436/524, 422/56
• US Class Current: 435/25
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10S 435/805 Test papers

Abstract

Non-aqueous, polymeric reagent film compositions for use with analytical test devices of the dry chemistry type are described. The film compositions are prepared from an organic solution of a copolymer formed by interaction of at least two monomers wherein

(a) the first monomer is a hydroxylated acrylate of the general formula ##STR1## where R1 is hydrogen or, more preferably, methyl, and R2 is a hydroxyalkyl group having from 1 to 5 carbon atoms, and

(b) the second monomer is a neutral acrylate of the general formula ##STR2## where R1 is hydrogen or, more preferably, methyl, and R3 is a substituted or unsubstituted alkyl group having from 1 to 8 carbon atoms.

Preferably, a third monomer is also included which is an amine-containing acrylate of the general formula ##STR3## where R1 is hydrogen or, more preferably, methyl, and R4 is a substituted or unsubstituted aminoalkyl or glycidyl group having from 1 to 5 carbon atoms or a surfactant polyethylene glycol group having from 10 to 30 carbon atoms. The film further comprises at least one enzyme in powder form dispersed therein and, optionally, a chromogenic indicator. The film composition is especially useful for determining components in body fluids, for example, for detecting glucose in whole blood, by rapid techniques.

Claims

1. A hydrophilic reagent composition for determining an analyte in a sample comprising:

(a) a nonaqueous copolymer formed from a mixture comprising;

(i) a first monomer of the general formula ##STR7## where R1 is hydrogen or methyl, and R2 is a hydroxyalkyl group having from 1 to 5 carbon atoms,

(ii) a second monomer of the general formula ##STR8## where R1 is hydrogen or methyl and R3 is a substituted or unsubstituted alkyl group having from 1 to 8 carbon atoms and wherein said second monomer is neutral in charge,

(iii) an organic solvent, and

(iv) an initiator in an amount sufficient for causing polymerization of the monomers;

(b) an enzyme in powder form homogeneously dispersed in said copolymer and reactive with said analyte; and

(c) an indicator in an amount sufficient to generate a detectable signal when in the presence of said analyte and said enzyme.

2. The composition of claim 1, wherein said copolymer is formed from a mixture further comprising a third monomer of the formula ##STR9## where R1 is hydrogen or methyl and R4 is hydrogen, an aminoalkyl or glycidyl group having from 1 to 5 carbon atoms or a surfactant polyethylene glycol group having from 10 to 30 carbon atoms, and wherein said third monomer is present in an amount less than about 5 percent by weight of the copolymer composition.

3. The composition of claim 1 or 2 wherein said first monomer is present in an amount of at least about 33 percent by weight of the copolymer composition.

4. The composition of claim 1 or 2 wherein said first monomer is present in an amount greater than about 50 percent by weight of the copolymer composition.

5. The composition of claim 1 or 2 wherein said first monomer is selected from the group consisting of hydroxyethyl methacrylate and hydroxypropyl methacrylate.

6. The composition of claim 1 or 2 wherein said second monomer is selected from the group consisting of butyl methacrylate and methyl methacrylate.

7. The composition of claim 2 wherein said third monomer is selected from the group consisting of dimethylaminoethyl methacrylate and t-butylaminoethyl methacrylate.

8. The composition of claim 1 or 2 wherein said organic solvent is comprised of a mixture of xylene and 1-methoxy-2-propanol.

9. The composition of claim 1 or 2 wherein said copolymer is nonionic.

10. The composition of claim 1 or 2 wherein said enzyme is selected from the group consisting of glucose oxidase and a peroxidase.

11. The composition of claim 1 or 2 wherein said indicator is selected from the group consisting of 3,3'"'"',5,5'"'"'-tetramethylbenzidine, a derivative thereof and an analog thereof.

12. The composition of claim 1 or 2, further comprising a ranging compound selected from the group consisting of 3-amino-9-(-aminopropyl)-carbozol dihydrochloride and butylated hydroxytoluene, wherein said ranging compound is present in an amount sufficient to reduce signal generation by said indicator, thereby making said composition useful for determining high levels of said analyte.

13. The composition of claim 1 or 2, further comprising a detergent selected from the group consisting of sodium dodecyl sulfate and sodium dodecylbenzene sulfonate.

14. The composition of claim 1 or 2, further comprising ground micro mica in an amount sufficient to reduce film tack.

15. An apparatus for determining an analyte in a sample comprising a carrier which is inert with respect to said analyte and a hydrophilic reagent composition coated thereon, said reagent composition comprising:

(a) a nonaqueous copolymer formed from a mixture comprising;

(i) a first monomer of the formula ##STR10## where R1 is hydrogen and methyl, and R2 is a hydroxyalkyl group having from 1 to 5 carbon atoms,

(ii) a second monomer of the formula ##STR11## where R1 is hydrogen or methyl and R3 is a substituted or unsubstituted alkyl group having from 1 to 8 carbon atoms and wherein said second monomer is neutral in charge,

(iii) an organic solvent, and

(iv) an initiator in an amount sufficient for causing polymerization of the monomers;

(b) an enzyme in powder form homogeneously dispersed in said copolymer and reactive with said analyte; and

(c) an indicator in an amount sufficient to generate a detectable signal when in the presence of said analyte and said enzyme.

16. The apparatus of claim 15, wherein said copolymer is formed from a mixture further comprising a third monomer of the formula ##STR12## where R1 is hydrogen and methyl and R4 is hydrogen, an aminoalkyl or glycidyl group having from 1 to 5 carbon atoms or a surfactant polyethylene glycol group having from 10 to 30 carbon atoms, and wherein said third monomer is selected from the group consisting of present in an amount less than about 5 percent by weight of the copolymer composition.

17. The apparatus of claim 15 or 16 wherein said first monomer is present in an amount of at least about 33 percent by weight of the copolymer composition.

18. The apparatus of claim 15 or 16 wherein said first monomer is present in an amount from about 10 to 70 percent by weight of the copolymer composition.

19. The apparatus of claim 15 or 16 wherein said first monomer is selected from the group consisting of hydroxyethyl methacrylate and hydroxypropyl methacrylate.

20. The apparatus of claim 15 or 16 wherein said second monomer is selected from the group consisting of butyl methacrylate and methyl methacrylate.

21. The apparatus of claim 16 wherein said third monomer is selected from the group consisting of dimethylaminoethyl methacrylate and t-butylaminoethyl methacrylate.

22. The apparatus of claim 15 or 16 wherein said organic solvent is comprised of a mixture of xylene and 1-methoxy-2-propanol.

23. The apparatus of claim 15 or 16 wherein said copolymer is nonionic.

24. The apparatus of claim 15 or 16 wherein said enzyme is selected from the group consisting of glucose oxidase and a peroxidase.

25. The apparatus of claim 15 or 16 wherein said indicator is selected from the group consisting of 3,3'"'"',5,5'"'"'-tetramethylbenzidine, a derivative thereof and an analog thereof.

26. The apparatus of claim 15 or 16, wherein said reagent composition further comprises a ranging compound selected from the group consisting of 3-amino-9-(-aminopropyl)-carbozol dihydrochloride and butylated hydroxytoluene, wherein said ranging compound is present in an amount sufficient to reduce signal generation by said indicator, thereby making said composition useful for determining high levels of said analyte.

27. The apparatus of claim 15 or 16, wherein said reagent composition further comprises a detergent selected from the group consisting of sodium dodecyl sulfate and sodium dodecylbenzene sulfonate.

28. The apparatus of claim 15 or 16, wherein said reagent composition further comprises ground micro mica in an amount sufficient to reduce film tack.

29. A method for determining an analyte in a sample by contacting said sample with a hydrophilic reagent composition to form a reaction mixture and observing said reaction mixture for the appearance of a detectable signal, wherein said reagent composition comprises:

(a) a nonaqueous copolymer formed from a mixture of;

(i) a first monomer of the formula ##STR13## where R1 is hydrogen or methyl, and R2 is a hydroxyalkyl group having from 1 to 5 carbon atoms,

(ii) a second monomer of the formula ##STR14## where R1 is hydrogen and methyl and R3 is a substituted or unsubstituted alkyl group having from 1 to 8 carbon atoms and wherein said second monomer is neutral in charge,

(iii) an organic solvent, and

(iv) an initiator in an amount sufficient for causing polymerization of the monomers;

(b) an enzyme in powder form homogeneously dispersed in said copolymer and reactive with said analyte; and

(c) an indicator in an amount sufficient to generate said detectable signal when in the presence of said analyte and said enzyme.

30. The method of claim 29, wherein said copolymer is formed from a mixture further comprises a third monomer of the general formula ##STR15## where R1 is hydrogen and methyl and R4 is hydrogen, an aminoalkyl or glycidyl group having from 1 to 5 carbon atoms or a surfactant polyethylene glycol group having from 10 to 30 carbon atoms, and wherein said third monomer is selected from the group consisting of present in an amount less than about 5 percent by weight of the copolymer composition.

31. The method of claim 29 or 30 wherein said first monomer is present in an amount of at least about 33 percent by weight of the copolymer composition.

32. The method of claim 29 or 30 wherein said first monomer is from about 10 to 70 percent by weight of the copolymer composition.

33. The method of claim 29 or 30 wherein said first monomer is selected from the group consisting of hydroxyethyl methacrylate and hydroxypropyl methacrylate.

34. The method of claim 29 or 30 wherein said second monomer is selected from the group consisting of butyl methacrylate and methyl methacrylate.

35. The method of claim 30 wherein said third monomer is selected from the group consisting of dimethylaminoethyl methacrylate and t-butylaminoethyl methacrylate.

36. The method of claim 29 or 30 wherein said organic solvent is comprised of a mixture of xylene and 1-methoxy-2-propanol.

37. The method of claim 29 or 30 wherein said copolymer is nonionic.

38. The method of claim 29 or 30 wherein said enzyme is selected from the group consisting of glucose oxidase or a peroxidase.

39. The method of claim 29 or 30 wherein said indicator is selected from the group consisting of 3,3'"'"',5,5'"'"'-tetramethylbenzidine, a derivative thereof and an analog thereof.

40. The method of claim 29 or 30, wherein said reagent composition further comprises a ranging compound selected from the group consisting of 3-amino-9-(-aminopropyl)-carbozol dihydrochloride and butylated hydroxytoluene, wherein said ranging compound is present in an amount sufficient to reduce signal generation by said indicator, thereby making said composition useful for determining high levels of said analyte.

41. The method of claim 29 or 30, wherein said reagent composition further comprises a detergent selected from the group consisting of sodium dodecyl sulfate and sodium dodecylbenzene sulfonate.

42. The method of claim 29 or 30, wherein said reagent composition further comprises ground micro mica in an amount sufficient to reduce film tack.