EP0048090A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Substrate specific galactose oxidase enzyme electrodes
Assignee: Yellow Springs Instrument Co Inc
•
Filing: 1981-08-19
•
Granted: —
•
Expires: 2001-08-19
Abstract
Abstract The relative specificity of the enzyme galactose oxidase for various substrates is controlled as a function of the electrical potential applied to the enzyme. The enzyme is incorporated into a thin layer, electrochemical cell laminate (40) having exterior membrane layers (42-44,42'-44') and an interior enzyme layer (46). An electrode (48) located within the enzyme layer applies varying electrical potentials to the enzyme. An intermediate electron transfer agent may be used to transfer electrons to and from the enzyme and electrode.
Inventors
- 👤 Jay Michael Johnson
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 10:44
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/001; C12Q1/004; Y; Y10; Y10S; Y10S435/00; Y10S435/817
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
143 words
A thin layer electrochemical cell for use in controlling the relative specificity of an enzyme for various substrates characterised by: a first outer membrane layer (34 or 42-44) adapted to be adjacent a substrate solution for allowing passage of low molecular weight materials into said cell (20 or 40) but excluding high molecular weight materials contained in said substrate solution, a second inner membrane layer (32 or 42'-44') adapted to be adjacent to a polarographic cell (10) for excluding passage of interfering low molecular weight materials and any mediator but permitting the passage of the electroactive reactant or product of the reaction between said substrate or substrates and an enzyme, an enzyme preparation (36 or 46) positioned between said first and second membrane layers, and electrode means (38 or 48) contacting said enzyme preparation for varying the electrical potential of said enzyme preparation.
Claim 2
Dependent Claim
143 words
A thin layer electrochemical cell for use in controlling the relative specificity of an enzyme for various substrates characterised by: a first outer membrane layer (34 or 42-44) adapted to be adjacent a substrate solution for allowing passage of low molecular weight materials into said cell (20 or 40) but excluding high molecular weight materials contained in said substrate solution, a second inner membrane layer (32 or 42'-44') adapted to be adjacent to a polarographic cell (10) for excluding passage of interfering low molecular weight materials and any mediator but permitting the passage of the electroactive reactant or product of the reaction between said substrate or substrates and an enzyme, an enzyme preparation (36 or 46) positioned between said first and second membrane layers, and electrode means (38 or 48) contacting said enzyme preparation for varying the electrical potential of said enzyme preparation.
Claim 3
Dependent Claim
14 words
The electrochemical cell of claim 1, in which said enzyme preparation contains galactose oxidase.
Claim 4
Dependent Claim
21 words
The electrochemical cell of claim 1 or 2, in which said first membrane layer (34 or 42-44) is a polycarbonate film.
Claim 5
Dependent Claim
35 words
The electrochemical cell of claim 1, 2 or 3,in which said second membrane layer (32 or 42'-44') is an essentially homogeneous material selected from the group consisting of silicone rubber, methyl methacrylate, and cellulose acetate.
Claim 6
Dependent Claim
19 words
The electrochemical cell of any preceding claim, in which said enzyme preparation additionally contains an intermediate electron transfer agent.
Claim 7
Dependent Claim
60 words
The electrochemical cell of any preceding claim, in which said first membrane layer (42-44) comprises a coupled membrane layer of a first polymeric material which acts as a support for a second polymeric material, said second polymeric material having a plurality of pores the diameter of which is less than the diameter of the pores in said first polymeric material.
Claim 8
Dependent Claim
62 words
The electrochemical cell of claim 6, in which said second membrane layer comprises a coupled membrane layer of a first polymeric material (44) which acts as a support for a second polymeric material (42), said second polymeric material (42) having a plurality of pores the diameter of which is less than the diameter of the pores in said first polymeric material (44).
Claim 9
Dependent Claim
29 words
A cell according to any preceding claim, in which said electrode means (38) comprises a layer of gold deposited on the back wall of said first membrane layer (34).
Claim 10
Dependent Claim
22 words
The electrochemical cell of any of claims 1 to 8, in which said electrode means (48) comprises a grid of gold wires.
Claim 11
Independent Claim
82 words
A method for quantitatively determining the amount of a particular substrate catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product characterised by the steps of: applying an electrical potential to said enzyme, said electrical potential being sufficient to control the redox potential of said multisubstrate enzyme, contacting said enzyme with a solution containing at least said particular substrate, and measuring for an electroactive reactant or product of the reaction mechanism.
Claim 12
Independent Claim
82 words
A method for quantitatively determining the amount of a particular substrate catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product characterised by the steps of: applying an electrical potential to said enzyme, said electrical potential being sufficient to control the redox potential of said multisubstrate enzyme, contacting said enzyme with a solution containing at least said particular substrate, and measuring for an electroactive reactant or product of the reaction mechanism.
Claim 13
Independent Claim
94 words
A method for quantitatively determining the amount of a particular substrate or substrates catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product, characterised by the steps of: applying at least two different electrical potentials to said enzyme, said electrical potentials being sufficient to render different the relative sensitivity of the enzyme for at least two of the substrates present, contacting said enzyme with a solution containing at least two substrates, and measuring for an electroactive reactant or product of the reaction mechanism.
Claim 14
Independent Claim
94 words
A method for quantitatively determining the amount of a particular substrate or substrates catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product, characterised by the steps of: applying at least two different electrical potentials to said enzyme, said electrical potentials being sufficient to render different the relative sensitivity of the enzyme for at least two of the substrates present, contacting said enzyme with a solution containing at least two substrates, and measuring for an electroactive reactant or product of the reaction mechanism.
Claim 15
Independent Claim
83 words
A method of measuring for the presence of substrates catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product,characterised by the steps of applying an electrical potential to said enzyme, contacting said enzyme with a solution containing at least one substrate catalyzed by said enzyme, varying the electrical potential applied and measuring for an electroactive reactant or product of the reaction mechanism as a function of the potential applied to said enzyme.
Claim 16
Independent Claim
83 words
A method of measuring for the presence of substrates catalyzed by a multisubstrate enzyme having a redox potential to react with oxygen and form hydrogen peroxide as a reaction product,characterised by the steps of applying an electrical potential to said enzyme, contacting said enzyme with a solution containing at least one substrate catalyzed by said enzyme, varying the electrical potential applied and measuring for an electroactive reactant or product of the reaction mechanism as a function of the potential applied to said enzyme.
Claim 17
Dependent Claim
17 words
The method of claim 10, 11 or 12, wherein hydrogen peroxide is the electroactive product being measured.
Claim 18
Dependent Claim
19 words
The method of claim 10, 11 or 12, wherein galactose oxidase is the multisubstrate enzyme having a redox potential.
Claim 19
Dependent Claim
185 words
A polarographic cell structure for use in polarographic analysis of a substrate in a substrate solution comprising a polarographic electrode means (16, 17, 18) mounted within said polarographic cell structure (10), said polarographic electrode means including means defining an active exposed working face, characterised by: a thin layer electrochemical cell (20 or 40) separating said polarographic electrode means(16, 17, 18) from said substrate solution, said thin layer electrochemical cell having, a first outer membrane layer (34 or 42-44) adapted to be adjacent the substrate solution for allowing passage of low molecular weight materials into said cell but excluding high molecular weight materials contained in said substrate solution, a second inner membrane layer (32 or 42'-44') adapted to be adjacent to the polarographic cell (10) for excluding passage of interfering low molecular weight materials but permitting the passage of the electroactive reactant or product of the reaction between said substrate(s) and an enzyme, an enzyme preparation (36 or 46) positioned between said first and second membrane layers, and electrode means (38 or 48) contacting said enzyme preparation for varying the electrical potential of said enzyme preparation.
Claim 20
Dependent Claim
185 words
A polarographic cell structure for use in polarographic analysis of a substrate in a substrate solution comprising a polarographic electrode means (16, 17, 18) mounted within said polarographic cell structure (10), said polarographic electrode means including means defining an active exposed working face, characterised by: a thin layer electrochemical cell (20 or 40) separating said polarographic electrode means(16, 17, 18) from said substrate solution, said thin layer electrochemical cell having, a first outer membrane layer (34 or 42-44) adapted to be adjacent the substrate solution for allowing passage of low molecular weight materials into said cell but excluding high molecular weight materials contained in said substrate solution, a second inner membrane layer (32 or 42'-44') adapted to be adjacent to the polarographic cell (10) for excluding passage of interfering low molecular weight materials but permitting the passage of the electroactive reactant or product of the reaction between said substrate(s) and an enzyme, an enzyme preparation (36 or 46) positioned between said first and second membrane layers, and electrode means (38 or 48) contacting said enzyme preparation for varying the electrical potential of said enzyme preparation.
Claim 21
Dependent Claim
14 words
The polarographic cell structure of claim 15, wherein said enzyme preparation contains galactose oxidase.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.