Enzymatic detection device for detecting a gaseous or aerosol substance

• US 5,504,006 A
• Filed: 12/11/1989
• Issued: 04/02/1996
• Est. Priority Date: 12/17/1988
• Status: Expired due to Fees

• Current Assignee: Dragerwerk AG
• Original Assignee: Dragerwerk AG
• Inventors: Rindt, Klaus-Peter, Scholtissek, Stephan
• Primary Examiner(s): Snay, Jeffrey R.
• Application Number:
  US07/448,235
• Time in Patent Office: 2,304 Days
• Field of Search: 435/807, 435/288, 435/291, 435/296, 436/167, 436/169, 436/181, 422/56, 422/58, 422/83, 422/85, 422/88, 422/86, 422/87
• US Class Current: 435/287.7
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: G01N 31/223 for investigating presence of specific gases or aerosols G01N31/221, G01N31/222 take precedence; actuation of fire alarm by presence of smoke or gases G08B17/10
• CPC Class Codes: Y10S 435/807 Gas detection apparatus

Abstract

The invention is directed to a detecting device for detecting gaseous substances and aerosols with the aid of an enzymatic reaction. This reaction takes place between a reactant and an enzyme which, on the one hand, is exposed to the substance to be detected and, on the other hand, is held by a carrier which in the form of a porous holder is in contact with a solution activating and maintaining the enzymatic reaction. The properties of the solution are changed by the enzymatic reaction and are evaluated. The detection device is so improved that a capability for evaluation is made possible which is quantitative and which is specific for a particular substance to be detected. The substance to be detected as a reactant can be stoichiometrically transferrable into a reaction product for quantitatively determining the substance to be detected by means of an enzymatic reaction.

Claims

1. A detection device for detecting a specific gaseous or aerosol substance defining an enzyme substrate, the detection device comprising:

a vessel defining an interior;

an enzyme;

a holder holding said enzyme and having an upper surface exposed to said substance;

an aqueous buffer solution held in said interior of said vessel;

a chromogen reagent disposed in said holder;

said holder having a lower surface immersed in said solution and being porous to facilitate the capillary transport of said solution into and along said holder to bring said enzyme to said upper surface where said solution prepares said enzyme to act as a catalyzer for facilitating a reaction involving at least said enzyme substrate and said chromogen reagent to thereby form a product having a coloration with an intensity dependent upon the quantity of the substance detected;

said chromogen reagent being disposed in said holder in lyophilic form below said upper surface.

2. A detection device for detecting a specific gaseous or aerosol substance defining an enzyme substrate, the detection device comprising:

a vessel defining an interior;

an enzyme;

a holder holding said enzyme and having an upper surface exposed to said substance;

an aqueous buffer solution held in said interior of said vessel and containing a chromogen reagent;

said holder having a lower surface immersed in said solution and being porous to facilitate the capillary transport of said solution into and along said holder to bring said enzyme to said upper surface where said solution prepares said enzyme to act as a catalyzer for facilitating a reaction involving at least said enzyme substrate and said chromogen reagent to thereby form a product having a coloration with an intensity dependent upon the quantity of the substance detected.

3. The detection device of claim 2, said chromogen reagent being present in said solution and said solution further containing at least one additional substance in the form of a cofactor reagent which together form a coloring after said reaction is completed.

4. The detection device of claim 2, said porous holder having an end portion facing toward the substance to be detected and said end portion accommodating said enzyme.

5. The detection device of claim 2, comprising a porous diffusion membrane for covering said porous holder with respect to the ambient containing said substance.

6. The detection device of claim 2, wherein the substance to be detected is hydrogen peroxide and the enzyme is horseradish peroxidase for which said solution contains 1 mMol/l 4-aminoantipyrine and 1 mMol of N-ethyl-N-sulphopropyl-m-toluidine in 50 mMol/l phosphate buffer pH-value 7.3 as coloring reagents and 2 Units peroxidase per ml.

7. The detection device of claim 2, wherein the substance to be detected is hydrogen peroxide for which said solution contains 2 Units (international units) of the enzyme horseradish peroxidase per ml and as a coloring reagent one mMol/l 2.2-acino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) in 0.1 Mol/l of TRIS-HCl-buffer pH 8.0.

8. A detection device for detecting a specific gaseous or aerosol substance defining an enzyme substrate, the detection device comprising:

a vessel defining an interior;

an enzyme;

a holder having an upper surface;

an aqueous buffer solution held in said interior of said vessel and containing a chromogen reagent;

a porous membrane covering said upper surface and exposed to said substance to be detected;

said enzyme being bound to said porous membrane;

said holder having a lower end communicating with said solution and said upper surface facing away from said solution and toward said membrane;

said holder being porous to facilitate the capillary transport of said solution into and along said holder to bring said solution into contact with said membrane to activate said enzyme so as to permit said enzyme to act as a catalyzer for facilitating a reaction involving at least said enzyme substrate and said chromogen reagent to thereby form a product having a coloration having an intensity dependent upon the quantity of the substance detected.

9. The detection device of claim 8, said chromogen reagent being present in said solution and said solution further containing at least one additional substance in the form of a cofactor reagent which together form a coloring after said reaction is completed.

10. The detection device of claim 8, said reagents being accommodated in said holder in lyophilic form below said upper surface;

said interior defining a reservoir for containing solutizing agents for mobilizing the lyophilic reaction partners;

said enzyme being made capable of said enzymatic reaction after a connection is established between the porous holder and said reservoir.

11. The detection device of claim 8, comprising a porous diffusion membrane for covering said porous holder with respect to the ambient containing said substance.

12. The detection device of claim 8, wherein the substance to be detected is hydrogen peroxide and the enzyme is horseradish peroxidase for which said solution contains 1 mMol/l 4-aminoantipyrine and 1 mMol of N-ethyl-N-sulphopropyl-m-toluidine in 50 mMol/l phosphate buffer pH-value 7.3 as coloring reagents and 2 Units peroxidase per ml.

13. The detection device of claim 8, wherein the substance to be detected is hydrogen peroxide for which said solution contains 2 Units (international units) of the enzyme horseradish peroxidase per ml and as a coloring reagent one mMol/l 2.2-acino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) in 0.1 Mol/l of TRIS-HCl-buffer pH 8.0.