Dry multilayer analytical elements for assaying transaminases

• US 5,462,858 A
• Filed: 12/29/1993
• Issued: 10/31/1995
• Est. Priority Date: 12/29/1993
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Bale Oenick, Marsha D., Sutherland, John W. H., Eikenberry, Jon N., Detwiler, Richard L.
• Primary Examiner(s): Thibodeau, Paul J.
• Assistant Examiner(s): CHEN, VIVIAN
• Application Number:
  US08/175,013
• Time in Patent Office: 671 Days
• Field of Search: 435/16, 435/15, 435/25, 435/28, 435/805, 436/169, 428/327, 428/328, 428/478.2, 428/475.2, 428/331, 422/55, 422/56, 422/57
• US Class Current: 435/16
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: Y10S 435/805 Test papers

Abstract

A dry multilayer analytical element for assaying transaminases wherein the analytical element comprises:

a) a transaminase substrate;

b) reagents that generate pyruvate in the presence of a transaminase and the transaminase substrate;

c) pyruvate oxidase and cofactors for pyruvate oxidase; and

d) reagents that generate a color in proportion to a concentration of hydrogen peroxide;

and wherein the analytical element has, in the following order, a spreading layer, a reagent layer, and a support wherein

i) the spreading layer contains the transaminase substrate and a material selected from the group consisting of polymer beads, barium sulfate, titanium oxide, and mixtures thereof;

ii) the reagent layer contains the pyruvate oxidase; and

iii) both a hardened gelatin layer and a subbing layer separate the reagent layer from the spreading layer.

Claims

1. A dry multilayer analytical element for assaying transaminases wherein said element comprises:

a) a transaminase substrate;

b) reagents that generate pyruvate in the presence of a transaminase and the transaminase substrate;

c) pyruvate oxidase and cofactors for pyruvate oxidase; and

d) reagents that generate a color in proportion to a concentration of hydrogen peroxide;

and wherein said element has, in the following order, a spreading layer, a reagent layer, and a support wherein

i) the spreading layer contains the transaminase substrate and a material selected from the group consisting of polymer beads, barium sulfate, titanium oxide, and mixtures thereof;

ii) the reagent layer contains the pyruvate oxidase; and

iii) both a hardened gelatin layer and a subbing layer separate the reagent layer from the spreading layer.

2. The element of claim 1 wherein the subbing layer comprises polyvinylpyrrolidone.

3. The element of claim 2 wherein the polyvinylpyrrolidone has molecular weight of 10,000 to 360,000.

4. The element of claim 3 wherein the polyvinylpyrrolidone has molecular weight of about 40,000.

5. The element of claim 1 or 4 wherein the reagent layer comprises from about 1000 to 4500 U/m² of pyruvate oxidase.

6. The element of claim 1 or 4 wherein the reagents that generate a color in proportion to the concentration of hydrogen peroxide are a leuco dye and horseradish peroxidase;

14. An element according to claim 1, 7 or 8 wherein the spreading layer comprises barium sulfate or titanium dioxide.

7. A dry multilayer analytical element for assaying aspartate aminotransaminase wherein said element comprises:

a) aspartate;

b) reagents that generate pyruvate in the presence of aspartate aminotransaminase and aspartate;

c) pyruvate oxidase and cofactors for pyruvate oxidase; and

d) reagents that generate a color in proportion to a concentration of hydrogen peroxide;

and wherein said element has, in the following order, a spreading layer, a reagent layer, and a support wherein

i) the spreading layer contains aspartate and a material selected from the group consisting of polymer beads, barium sulfate, titanium oxide, and mixtures thereof;

ii) the reagent layer contains the pyruvate oxidase; and

iii) both a hardened gelatin layer and a subbing layer separate the reagent layer from the spreading layer.

9. The element of claim 7 or 8 wherein the subbing layer is polyvinylpyrrolidone.

10. The element of claim 9 wherein the polyvinylpyrrolidone has a molecular weight of 10,000 to 360,000.

11. The element of claim 10 wherein the polyvinylpyrrolidone has a molecular weight of about 40,000.

12. The element of claim 7 or 8 wherein the reagent layer comprises from about 1000 to 4500 U/m² of pyruvate oxidase.

13. The element of claim 7 or 8 wherein the reagents that generate a color in proportion to the concentration of hydrogen peroxide are a leuco dye and horseradish peroxidase.

8. A dry multilayer analytical element for assaying alanine aminotransaminase wherein said element comprises:

a) alanine;

b) reagents that generate pyruvate in the presence of alanine aminotransaminase and alanine;

c) pyruvate oxidase and cofactors for pyruvate oxidase; and

d) reagents that generate a color in proportion to a concentration of hydrogen peroxide;

and wherein said element has, in the following order, a spreading layer, a reagent layer, and a support wherein

i) the spreading layer contains alanine and a material selected from the group consisting of polymer beads, barium sulfate, titanium oxide, and mixtures thereof;

ii) the reagent layer contains the pyruvate oxidase; and

iii) both a hardened gelatin layer and a subbing layer separate the reagent layer from the spreading layer.