Analytical element containing photosensitive compound and filter layer and method of use

• US 4,788,140 A
• Filed: 02/18/1986
• Issued: 11/29/1988
• Est. Priority Date: 02/18/1986
• Status: Expired due to Fees

• Current Assignee: Biogen, Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Wellman, Ronald A., Shafer, Mark E., Burdick, Brent A., Findlay, John B.
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US06/830,036
• Time in Patent Office: 1,015 Days
• Field of Search: 435/7, 435/17, 435/28, 435/805, 422/56
• US Class Current: 435/17
• CPC Class Codes: C12Q 1/50 involving creatine phosphokinase
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10S 435/805 Test papers

Abstract

A multilayer analytical element for the determination of a clinically significant enzyme analyte comprises a photosensitive compound (e.g. a photosensitive dye or dye precursor) and a filter layer containing one or more filter dyes. The filter layer is situated in the element such that incident radiation used to detect a density change resulting from interaction of the analyte and the photosensitive compound passes through the filter layer before it reaches the photosensitive compound. The use of the filter layer inhibits premature changes in the photosensitive compound caused by incident radiation. This element is particularly useful for the determination of creatine kinase or one of its isoenzymes, e.g. creatine kinase-MB.

Claims

1. A self-supporting analytical element comprising:

a layer comprising an absorbent carrier material containing an interactive composition for a clinically significant enzyme analyte, said composition comprising a photosensitive compound capable of providing a detectable optical density change at a predetermined characteristic wavelength in response to reaction of the analyte with its substrate, and

a filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said characteristic wavelength produced in response to a reaction of the analyte with its substrate.

2. The element of claim 1 wherein said photosensitive compound is a photosensitive dye or dye precursor.

3. The element of claim 2 wherein said photosensitive dye precursor is an imidazole leuco dye.

4. A multilayer analytical element comprising

a nonporous support having thereon two or more layers, and contained in at least one of said layers, an interactive composition for a clinically significant enzyme analyte, said composition comprising a photosensitive dye or dye precursor capable of providing a detectable optical density change at a predetermined characteristic wavelength in response to action of said analyte on a substrate,

at least one of said layers being a porous spreading layer, and a second being a filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said predetermined characteristic wavelength produced in response to a reaction of the analyte with the substrate.

5. The element of claim 4 wherein said support is transparent to incident radiation and said photosensitive dye or dye precursor is in a layer located on the opposite side of said support from said filter layer.

6. The element of claim 4 wherein said filter layer is located between said support and a layer containing said photosensitive dye or dye precursor.

7. The element of claim 4 wherein said photosensitive dye precursor is an imidazole leuco dye.

8. A multilayer analytical element for the determination of creatine kinase or an isoenzyme thereof comprising a nonporous transparent support having thereon three or more layers including a filter layer, and contained in at least one of said layers other than said filter layer, an interactive composition for creatine kinase, said composition comprising one or more reagents which cumulatively provide a detectable optical density change at a predetermined characteristic wavelength in response to action of creatine kinase on a substrate, one of said reagents being a photosensitive dye or dye precursor,

said support having on one side:

said filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said predetermined characteristic wavelength, produced in response to a reaction of creatine kinase or an isoenzyme thereof with its substrate and

on the other side, in order, two or more layers including;

a layer containing said photosensitive dye or dye precursor, and

an outermost porous spreading layer.

9. The element of claim 8 wherein said photosensitive dye precursor is a triarylimidazole leuco dye.

10. The element of claim 8 wherein said interactive composition further comprises the reagents creatine phosphate, adenosine-5'"'"'-diphosphate, glycerol, glycerol kinase and α

-glycerophosphate oxidase, which reagents are located in any of said layers other than said filter layer.

11. The element of claim 8 further comprising creatine kinase-M antibodies immobilized in said porous spreading layer.

12. The element of claim 8 wherein said filter layer contains filter dyes cumulatively capable of filtering incident radiation having a wavelength of about 500 nm or less.

13. A method for the determination of a clinically significant enzyme analyte comprising the steps of:

A. contacting a sample of a liquid suspected of containing a clinically significant enzyme analyte with an analytical element comprising;

a layer comprising an absorbent carrier material containing a substrate and an interactive composition for said analyte, said composition comprising a photosensitive compound which is capable of providing an optical density change at a predetermined characteristic wavelength in response to action of said analyte on said substrate, and

a filter layer comprising at least one filter dye selected to absorb incident radiation other than radiation at said characteristic wavelength,

to produce an optical density change detectable at said predetermined characteristic wavelength as a result of the presence of said enzyme in said sample, and

B. determining said optical density change at said predetermined characteristic wavelength by directing incident radiation from transmission, reflection or fluorescent spectrophotometry to said element in such fashion that said incident radiation passes through said filter layer before it reaches said absorbent layer.

14. A method for the determination of total creatine kinase or an isoenzyme thereof comprising the steps of:

A. contacting a sample of a liquid suspected of containing creatine kinase or an isoenzyme thereof with a multilayer analytical element comprising a nonporous transparent support having thereon three or more layers including a filter layer, and contained in at least one of said layers other than said filter layer, a substrate and an interactive composition for creatine kinase, said composition comprising one or more reagents which cumulatively provide a detectable optical density change at a predetermined characteristic wavelength in response to action of creatine kinase on said substrate, one of said reagents being a photosensitive dye or dye precursor,

said support having on one side;

said filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said predetermined characteristic wavelength, produced in response to a reaction of creatine kinase or an isoenzyme thereof with its substrate and

on the other side, in order, two or more layers including;

a layer containing said photosensitive dye or dye precursor, and

an outermost porous spreading layer, to produce an optical density change detectable at said predetermined characteristic wavelength as a result of the presence of creatine kinase or an isoenzyme thereof in said sample, and

B. determining said optical density change at said predetermined characteristic wavelength by directing incident radiation from transmission or reflection spectrophotometry to said element in such fashion that said incident radiation passes through said filter layer before it reaches said photosensitive dye or dye precursor layer.

15. The method of claim 14 wherein said filter layer comprises one or more filter dyes which are capable of filtering incident radiation having a wavelength of about 500 nm or less and said detectable optical density change is determined at a predetermined characteristic wavelength of greater than 500 nm.

16. The method of claim 14 for the determination of creatine kinase-MB wherein said element comprises creatine kinase-M antibodies immobilized in its porous spreading layer.

17. The method of claim 14 wherein said photosensitive dye is a triarylimidiazole leuco dye and said interactive composition comprises adenosine-5'"'"'-diphosphate, glycerol, glycerol kinase and α

-glycerophosphate oxidase.

18. The method of claim 17 wherein said triarylimidazole leuco dye is 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-bis(4-dimethylaminophenyl)imidazole.

19. A multilayer analytical element for the determination of creatine kinase or an isoenzyme thereof comprising a nonporous transparent support having thereon three or more layers including a filter layer, and contained in at least one of said layers other than said filter layer, an interactive composition for creatine kinase, said composition comprising one or more reagents which cumulatively provide a detectable optical density change at a predetermined characteristic wavelength in response to action of creation kinase on a substrate, one of said reagents being a photosensitive dye or dye precursor,

said support having on one side, in order, three or more layers including:

said filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said predetermined characteristic wavelength, produced in response to a reaction of creatine kinase or an isoenzyme thereof with its substrate and

a layer containing said photosensitive dye or dye precursor, and

an outermost porous spreading layer.

20. A multilayer analytical element for the determination of creatine kinase or an isoenzyme thereof comprising a nonporous support having thereon three or more layers including a filter layer, and contained in at least one of said layers other than said filter layer, an interactive composition for creatine kinase, said composition comprising one or more reagents which cumulatively provide a detectable optical density change at a predetermined characteristic wavelength in response to action of creatine kinase on a substrate, one of said reagents being a photosensitive dye or dye precursor,

said support having on one side, in order, three or more layers including:

a layer containing said photosensitive dye or dye precursor,

said filter layer containing at least one filter dye selected to absorb incident radiation other than radiation at said predetermined characteristic wavelength, produced in response to a reaction of creatine kinase or an isoenzyme thereof with its substrate and

an outermost porous spreading layer.