Multilayer reagent test strips and methods for using the same to quantify glycated protein in a physiological sample

• US 20030211564A1
• Filed: 05/10/2002
• Published: 11/13/2003
• Est. Priority Date: 05/10/2002
• Status: Active Grant

• Current Assignee: Cilag Gmbh International (Johnson & Johnson)
• Original Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Inventors: Guo, Sherry, Leong, Koon-Wah, Qian, Suyue
• Granted Patent: US 6,951,728 B2
• Time in Patent Office: Days
• US Class Current: 435/23
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

Multilayer reagent test strips for quantitating glycated protein in a fluid sample, as well as methods for using the same, are provided. The subject multilayer test strips include at least a filter layer, a proteinase layer and a ketoamine oxidase signal producing and fluid flow control system layer. In using the subject test strips, a fluid sample is applied to the test strip and a signal is generated that can be employed to quantitate the glycated protein level in the sample. The quantitated glycated protein level can then be employed to determine the amount of glycated protein in the fluid sample. Also provided are kits and systems that include the subject test strips and find use in practicing the subject methods. The subject compositions and methods find use in glycated protein monitoring applications, among other utilities.

Claims

1. A multilayer test strip comprising:

(a) a blood separation element for separating red blood cells from plasma;

(b) a protease layer in fluid communication with said blood separation element; and

(c) a ketoamine oxidase signal producing system layer in fluid communication with said protease layer.

2. The multilayer test strip according to claim 1, wherein ketoamine oxidase signal producing system layer is hydrophobic.

3. The multilayer test strip according to claim 2, wherein said layers are present in a stacked configuration.

4. The multilayer test strip according to claim 3, wherein said stacked layers are present in a chamber of a support element bounded by a substrate strip and a guard piece.

5. The multilayer test strip according to claim 4, wherein said strip has a configuration as shown in FIG. 1.

6. The multilayer test strip according to claim 1, wherein said protease layer comprises a porous matrix and a protease that cleaves proteins to produce ketoamine oxidase accessible ketoamine bonds.

7. The multilayer test strip according to claim 6, wherein said protease is protease XIV.

8. The multilayer test strip according to claim 1, wherein said ketoamine oxidase signal producing system layer comprises a porous matrix, a ketoamine oxidase, a peroxidase and at least one indicator compound.

9. The multilayer test strip according to claim 8, wherein said porous matrix of said ketoamine oxidase signal producing system layer is positively charged.

10. The multilayer test strip according to claim 9, wherein said positively charge porous matrix comprises nylon.

11. The multilayer test strip according to claim 8, wherein said indicator compound comprises a urea derivative dye.

12. The multilayer test strip according to claim 11, wherein said urea derivative dye is a negatively charged urea derivative dye.

13. The multilayer test strip according to claim 12, wherein said urea derivative dye has the formula:

R¹ R²NCONHR³, wherein R¹, R² taken together is a N, N-di-substituted aminoaryl and R³ is selected from the group consisting of carboxyalkyl, alkoxycarbonyl, alkylcarbonyl, arylsulfonyl, sulfoaryl and carboxyaryl.

14. The multilayer test strip according to claim 13, wherein said urea derivative dye is selected from group consisting of phenothiazine derivative dyes, phenoxazine derivative dyes and diphenylamine derivative dyes.

15. The multilayer test strip according to claim 14, wherein said urea derivative dye is 10-(carboxymethylaminocarbonyl)3,7-bis(dimethylamino)phenothiazine or a salt thereof.

16. The multilayer test strip according to claim 8, wherein said peroxidase is horseradish peroxidase.

17. A measurement system for measuring an amount of glycated protein in a fluid sample, said system comprising:

(a) a multilayer test strip according to claim 1;

and

(b) a signal detection instrument for detecting signal produced on said multilayer test strip.

18. A method for quantifying the amount of glycated protein in a physiological sample, said method comprising:

(a) applying said physiological sample to a multilayer test strip according to claim 1;

(b) detecting a signal produced on said test strip to quantify the amount of glycated protein in said physiological sample.

19. The method according to claim 18, wherein said physiological sample is whole blood.

20. The method according to claim 18, wherein said detecting step is performed by an automated instrument.

21. A kit for use in determining the concentration of glycated protein in a physiological sample, said kit comprising:

(a) a multilayer reagent test strip according to claim 1;

and

(b) at least one of;

(i) a means for obtaining a physiological sample and

(ii) a control.

22. The kit according to claim 21, wherein said means for obtaining said physiological sample is a lance.

23. The kit according to claim 21, wherein said control comprises a standardized concentration of glycated protein.

24. The kit according to claim 21, wherein said kit comprises both said means for obtaining said physiological sample and control.

25. A signal detection instrument having present therein a multilayer reagent test strip according to claim 1.