Multi-layer analytical element with preserved enzyme containing spreading layer

• US 4,914,020 A
• Filed: 01/31/1986
• Issued: 04/03/1990
• Est. Priority Date: 02/05/1985
• Status: Expired due to Fees

• Current Assignee: Konishiroku Photo Industry Co., Ltd., Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Konishiroku Photo Industry Co., Ltd., Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Koyama, Mikio, Kobayashi, Morio, Arai, Kazumi, Zanma, Souichi, Momose, Takasi, Furukawa, Kunihiro, Okaniwa, Kenichiro
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US06/824,450
• Time in Patent Office: 1,523 Days
• Field of Search: 422/56, 436/810, 435/4, 435/23, 435/24, 435/188
• US Class Current: 435/4
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements

Abstract

An analytical element to measure a specific component in a fluid samples comprising support, a layer containing a reagent provided the support and a spreading layer provided on above the reagent containing layer. In the spreading layer, an emzyme necessary for the reaction to produce a product capable of being detected with said reagent from the specific component is contained. The enzyme is protected from deterioration of activity during a manufacturing and preservation of the analytical element by means of the enzyme is included in the spread layer as a dispersion mixture with a protein and/or polypeptide. An accurate analytical result is stably obtained by the analytical element.

Claims

1. An analytical element for the analysis of a specific component in a fluid, said element comprising:

(a) a support,

(b) a layer containing a reagent provided on said support,

(c) a spreading layer provided on said reagent-containing layer, said spreading layer having a porous structure, and

(d) a dispersion mixture contained in the porous structure of said spreading layer and including a mixture of an enzyme and a protein and/or polypeptide, formed by freeze-drying a mixture of said enzyme and said protein and/or said polypeptide from an aqueous solution thereof, said enzyme being of a type supporting a reaction with said specific component to produce a product capable of being detected with said reagent, and said protein and/or polypeptide being substantially free from a substance disturbing said reaction or analysis whereby the protein and/or polypeptide prevent rapid deterioration of the enzyme.

2. The analytical element of claim 1 wherein said enzyme is a hydrolase.

3. The analytical element of claim 2 wherein said hydrolase is a cholesterolesterase or a lipoproteinlipase.

4. The analytical element of claim 1 wherein said enzyme is an oxidizing enzyme.

5. The analytical element of claim 4 wherein said oxidizing enzyme is selected from the group consisting of cholesteroloxydase, glucoseoxydase, peroxydase and uricase.

6. The analytical element of claim 1 wherein said enzyme is a dehydrase.

7. The analytical element of claim 6 wherein said dehydrase is a diaphorase or a glutamic acid dehydrase.

8. The analytical element of claim 1 wherein an amount of said enzyme contained in said spreading layer is from 10 U/m² to 100,000 U/m² of said layer.

9. The analytical element of claim 1 wherein said protein or polypeptide compound is selected from the group consisting of albumin, globulin, gelatin and decomposed gelatin.

10. The analytical element of claim 1 wherein a proportion of the enzyme to the protein and/or the polypeptide compound is from 10:

1 to 1;

5000 by weight.

11. The analytical element of claim 9 wherein the proportion of the enzyme to the protein and/or the polypeptide compound is from 1:

1 to 1;

200 by weight.

12. The analytical element of claim 1 wherein said spreading layer comprises fiborous material.

13. The analytical element of claim 1 wherein said spreading layer comprises combined particles.

14. The analytical element of claim 1 wherein the thickness of said spreading layer is from about 100 to about 500 μ

m.

15. The analytical element of claim 14 wherein the thickness of said spreading layer is from about 150 to about 350 μ

m.

16. The analytical element of claim 1 wherein the porosity of said spreading layer is from about 20 to about 50%.

17. The analytical element of claim 8 wherein said protein or polypeptide compound is selected from the group consisting of albumin, globulin, gelatin and decomposed gelatin.

18. The analytical element of claim 17 wherein a proportion of the enzyme to the protein and/or the polypeptide compound is from 10:

1 to 1;

5000 by weight.

19. The analytical element of claim 18 wherein the proportion of the enzyme to the protein and/or the polypeptide compound is from 1:

1 to 1;

200 by weight.

20. The analytical element of claim 19 wherein the thickness of said spreading layer is from about 100 to about 500 μ

m.

21. The analytical element of claim 20 wherein the thickness of said spreading layer is from about 150 to about 350 μ

m.

22. The analytical element of claim 21 wherein the porosity of said spreading layer is from about 20 to about 50%.

23. The analytical element of claim 17 wherein said enzyme is selected from the group consisting of hydrolase, oxidizing enzymes, and dehydrase.