Claims
- 1. A method for determining an analyte, said method comprising the steps of:
- A. forming a temporary laminate by overlaying a plate adapted for electrically induced migration and containing a plurality of analytes which have been electrically separated from one another, with a dry, transparent analytical element containing a water-insoluble binder material having pre-dispersed therein a reactive component which will react with at least one of said analytes to form a non-diffusible, detectable species solely in said element, and
- B. removing said analytical element from said plate and detecting said detectable species in said analytical element.
- 2. The method of claim 1 wherein said analytes are proteins.
- 3. The method of claim 1 wherein said interactive composition reacts to provide a non-diffusible chromogen.
- 4. The method of claim 1 wherein after step A and prior to step B, said plate-element laminate is incubated at a temperature up to about 45.degree. C. for a time sufficient for said analyte to be detected to react with said reactive component to generate a visible representation of the analyte.
- 5. A method for determining an analyte in an aqueous liquid, said method comprising the steps of:
- A. contacting an electrophoretic plate with a sample of said liquid,
- B. electrically separating a plurality of analytes from one another in said plate,
- C. forming a temporary laminate by overlaying said plate with a dry, transparent analytical element containing a water-insoluble binder material having predispersed therein an reactive component which will react with at least one of said analytes to form a non-diffusible, detectable species solely in said element, and
- D. removing said element from said plate and detecting said detectable species in said element.
- 6. The method of claim 5 wherein said reactive component reacts to provide a non-diffusible chromogen.
- 7. The method of claim 5 wherein said plurality of analytes are proteins or nucleic acids.
- 8. The method of claim 5 wherein said plurality of analytes are isoenzymes or enzymes.
- 9. The method of claim 8 wherein said plurality of isoenzymes are isoenzymes of creatine kinase, alkaline phosphotase or lactate dehydrogenase.
- 10. The method of claim 5 wherein said dry analytical element comprises a support having thereon at least one layer containing said binder material and said reactive component.
- 11. A method for detecting a plurality, (N), of analytes, said method comprising the steps of:
- A. forming a temporary laminate by overlaying a plate adapted for electrically induced migration and containing said plurality of analytes which have been separated from one another electrically, with a first analytical element containing a water-insoluble binder material having pre-dispersed therein a reactive component which will react with the first of said analytes while laminated to provide a first detectable species solely in said first element,
- B. removing said first element from said plate and
- C. repeating steps A and B up to N-1 times, using in each repetition of steps A and B, said plate and a respectively different dry analytical element containing a water-insoluble binder material having pre-dispersed therein a respective reactive component which will react with the respective analyte while laminated to form a respective detectable species solely in said respectively different element,
- provided that said reactive components of at least the first (N-1) detections react with said respective analytes to provide respective non-diffusible detectable species,
- and wherein each of said analytical elements is provided as a dry, transparent element.
- 12. The method of claim 11 wherein said plurality of analytes are proteins or nucleic acids.
- 13. The method of claim 12 wherein said plurality of proteins are enzymes or isoenzymes.
- 14. The method of claim 11 wherein after each performance of step A and before each performance of step B, said respective plate-element laminate is incubated at a temperature of up to about 45.degree. C. and for a time sufficient for said respective analyte to react with said respective reactive component to generate a visible representation of the analytes, and each of said elements is essentially transparent.
- 15. A method for the determination of creatine kinase isoenzymes, said method comprising the steps of:
- A. forming a temporary laminate by overlaying an electrophoretic plate which contains said isoenzymes which have been electrically separated from one another with a dry, transparent analytical element,
- said element comprising before said Step A, a nonporous support having thereon, in order and in fluid contact, a first reagent layer containing peroxidase, a-glycerophosphate oxidase and a leuco dye which is capable of providing a non-diffusible detectable dye in response to said isoenzymes solely in said element, and a second reagent layer containing a water-insoluble binder material having dispersed therein creatine phosphate, adenosine diphosphate, glycerol and glycerol kinase, and
- B. removing said element from said plate and examining said element to determine said detectable dye in said element.
- 16. The method of claim 15 wherein said electrophoretic plate is composed of agarose, polyacrylamide or acrylamide.
- 17. The method of claim 15 wherein said water-insoluble binder material is hardened gelatin.
- 18. A diagnostic kit for the determination of an analyte, said kit comprising:
- a plate adapted for electrically induced migration of said analyte, and
- a dry transparent analytical element comprising a water-insoluble binder material having pre-dispersed therein a reactive component which is capable of reacting with said analyte to form a non-diffusible detectable species solely in said element.
- 19. The kit of claim 18 wherein said element comprises a nonporous support having thereon a reagent layer comprising said binder material and said reactive component.
- 20. The kit of claim 18 wherein said reactive component is capable of reacting with said analyte to provide a non-diffusible chromogen.
- 21. A method for determining an analyte in an aqueous liquid, said method comprising the steps of:
- A. contacting an electrophoretic plate with a sample of said liquid;
- B. electrically separating a plurality of analytes from one another in said plate;
- C. before or after Step B, adding an enzyme to said plate containing the analytes, in a form that selectively binds the enzyme to one of said analytes;
- D. forming a laminate by overlaying said plate with a dry transparent analytical element comprising a binder and a reactive component predispersed in said binder which will react with at least said enzyme to provide a non-diffusible, detectable species solely in said element; and
- E. removing said element from said plate and detecting said detectable species in said element.
- 22. The method of claim 21, wherein said enzyme is bound to an antibody specific to said analyte or to an antibody to said analyte.
- 23. The method of claim 21, wherein said Step C occurs after said Step B.
- 24. The method of claim 10 wherein said element comprises at least a detection layer and a layer separate from said detection layer comprising an enzyme substrate, on said support.
- 25. A method for the detection of intracellular enzymes in live cells, comprising
- disposing such cells on a support,
- and overlaying the support with a dry, transparent element comprising a water-insoluble binder and a reactive component pre-dispersed in said binder and capable of reacting with at least one of said enzymes to produce a detectable change.
- said reactive component further including a surfactant of the type and in an amount effective to induce leakage of said at least one enzyme out of the cells.
- 26. A method as defined in claim 25, wherein the enzyme is creatine kinase and said surfactant is selected from the group consisting of a sodium salt of an alkylaryl polyether sulfonate and sodium alkyl naphthalene sulfonate.
- 27. A method as defined in claim 25, wherein the enzyme is alkaline phosphotase and said surfactant is an octylphenoxy polyethoxy ethanol.
- 28. A method as defined in claim 25, wherein the enzyme is lactate dehydrogenase and said surfactant is an octylphenoxy polyethoxy ethanol.
RELATED APPLICATIONS
This is a continuation-in-part of application Ser. No. 68,767 filed on June 29, 1987 which is a continuation-in-part of Ser. No. 811,031 filed Dec. 19, 1985, both now abandoned.
US Referenced Citations (5)
Foreign Referenced Citations (3)
Number |
Date |
Country |
1048910 |
Oct 1976 |
CAX |
1072428 |
Oct 1978 |
CAX |
230762 |
May 1987 |
EPX |
Non-Patent Literature Citations (3)
Entry |
Mickle et al, Clin. Chem. 24 (4), pp. 698-700 (1978) "Improved Technique for Developing Creatine Kinase Isoenzyme Bands by Using a Substrate in Gelatin Matrix". |
1984-1985 Enzyme Systems Products Expanding Catalog. |
Smith, J. Histochem. Cytochem., 32 (12), pp. 1265-1274 (1984) "Identification of Protease Isozymes after Analytical Isoelectric Focusing Using Fluorogenic Substrates Impregnated into Cellulose Membranes". |
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
68767 |
Jun 1987 |
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Parent |
811031 |
Dec 1985 |
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