Binder composition and analytical element having stabilized peroxidase in layer containing the composition

• US 5,155,024 A
• Filed: 07/10/1986
• Issued: 10/13/1992
• Est. Priority Date: 07/10/1986
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Eikenberry, Jon N.
• Primary Examiner(s): Kepplinger
• Assistant Examiner(s): Scheiner, Toni R.
• Application Number:
  US06/884,249
• Time in Patent Office: 2,287 Days
• Field of Search: 435/4, 435/7, 435/28, 435/805, 435/810, 435/291, 435/188, 422/56, 422/57, 422/58, 422/60, 422/61, 436/518, 436/808, 436/810, 436/7.92, 436/7.9, 436/25
• US Class Current: 435/7.9
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

An analytical element has a peroxidase-labeled ligand analog distributed within a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol). As a result, the peroxidase retains more of its stability prior to use. Such elements can be used to determine any of a number of immunologically reactive analytes, such as digoxin.

Claims

1. An analytical element comprising an absorbent carrier material, said element divided into two or more discrete zones, and containing in one of said zones a peroxidase-labeled ligand analog for an immunologically reactive ligand uniformly distributed in a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol).

2. The element of claim 1 further comprising an interactive composition which is capable of reacting with said ligand analog to provide a spectrophotometric signal in the presence of a substrate for peroxidase.

3. The element of claim 2 wherein said interactive composition comprises α

-glycerol phosphate oxidase and a triarylimidazole leuco dye.

4. The element of claim 2 further comprising in at least one of said zones an immobilized receptor for said immunologically reactive ligand.

5. The element of claim 1 wherein said binder composition comprises an additional binder material selected from the group consisting of gelatin, polyacrylic acid, acrylamide-N-vinyl-2-pyrrolidone copolymers, polyacrylamide, poly(N-vinyl-2-pyrrolidone) and water-absorbent starch-containing polymers.

6. The element of claim 1 wherein poly(vinyl alcohol) is present in the binder composition in an amount of at least about 80 percent, by weight.

7. A multilayer analytical element comprising a nonporous support having thereon, in order,

a registration layer,

a water-soluble layer containing a peroxidase-labeled ligand analog for an immuno-logically reactive ligand uniformly distributed in a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol), and

a porous spreading layer,

said element further comprising an interactive composition which is capable of interacting with said ligand analog to provide a spectrophotometric signal in the presence of a substrate for peroxidase.

8. The element of claim 7 wherein said ligand analog is peroxidase-labeled digoxin.

9. The element of claim 7 wherein said interactive composition comprises α

-glycerol phosphate oxidase and a triarylimidazole leuco dye.

10. The element of claim 7 further comprising an immobilized receptor for said immunologically reactive ligand in said spreading layer.

11. The element of claim 7 wherein said ligand analog is present in a coverage of at least about 10^-6 g/m².

12. A method for the determination of an immunologically reactive ligand comprising the steps of:

A. in the presence of a receptor for said ligand, contacting a sample of a liquid suspected of containing said ligand with an analytical element comprising an absorbent carrier material, said element divided into two or more discrete zones, and containing in one of said zones a peroxidase-labeled ligand analog for said ligand uniformly distributed in a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol), said contacting carried out in such a manner as to form a complex of receptor and ligand analog, and

B. determining the amount of said ligand as a result of the presence of complexed or uncomplexed ligand analog.

13. The method of claim 12 wherein said receptor for said ligand is immobilized in one of said zones of said element.

14. The method of claim 12 wherein said liquid sample is contacted with said element in such a manner that said receptor-ligand analog complex is immobilized in said element and horizontal separation of uncomplexed ligand analog from immobilized complex is effected.

15. The method of claim 14 wherein said separation is accomplished with a wash step subsequent to said contacting step.

16. The method of claim 12 wherein said element comprises a nonporous support having thereon, in order,

a registration layer,

a layer containing said ligand analog uniformly distributed in a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol), and

a porous spreading layer as said absorbent carrier material.

17. The method of claim 12 for the determination of digoxin in a biological fluid wherein said ligand analog is peroxidase-labeled digoxin.

18. The method of claim 12 wherein said complexed ligand analog is measured.

19. A multilayer analytical element comprising a nonporous support having thereon, in order,

a porous spreading layer, and

a water-soluble layer containing a peroxidase-labeled ligand analog for an immunologically reactive ligand uniformly distributed in a water-soluble binder composition comprising at least about 50 percent, by weight, of poly(vinyl alcohol).