Optical biosensor with enhanced activity retention for detection of halogenated organic compounds
First Claim
1. A distal tip of a biosensor ion sensing transducer for use in detecting an analyte comprising an halogenated organic compound in an environment, the tip comprising:
- a biocomponent comprising at least one enzyme selected from the group consisting of hydrolases, subclass EC 3.8, and lyases, subclass EC 4.5, for carrying out a dehalogenation of the compound, said biocomponent immobilized to a surface of the tip by means selected from the group consisting of (a) entrapment within a hydrogel;
(b) entrapment within a polymeric network;
(c) (micro)encapsulation;
(d) covalent-bonding;
and (e) adsorption;
a treatment of said biocomponent for maintaining a period of enzymatic efficacy; and
said biocomponent stabilized by means selected from the group consisting of crosslinking a surface of said immobilized biocomponent, crosslinking a polymer layer to said biocomponent, adding a gel-hardening agent to said biocomponent, adding a stabilizing agent to said biocomponent, and modifying a component of said immobilizing means.
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Accused Products
Abstract
A distal tip, and associated method of producing, of a biosensor ion sensing transducer for use in detecting an analyte comprising an halogenated organic compound in an environment such as soil or an aqueous environment. Features include: a biocomponent comprising at least one enzyme for carrying out a dehalogenation of the compound; the biocomponent is immobilized to a surface of the tip; a treatment of the biocomponent for maintaining a period of enzymatic efficacy; and the biocomponent stabilized by means preferably selected from the group consisting of crosslinking a surface of the immobilized biocomponent, crosslinking a polymer layer to the biocomponent, adding a gel-hardening agent to the biocomponent, adding a stabilizing agent to the biocomponent, and modifying a component of the immobilizing means. Immobilization can be carried out by: (a) entrapment within a hydrogel; (b) entrapment within a polymeric network; (c) (micro)encapsulation; (d) covalent-bonding; and (e) adsorption. The dehalogenase of the biocomponent may be selected from the group consisting of hydrolases, subclass EC 3.8, and lyases, subclass EC 4.5 as listed in Table 2A.
20 Citations
15 Claims
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1. A distal tip of a biosensor ion sensing transducer for use in detecting an analyte comprising an halogenated organic compound in an environment, the tip comprising:
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a biocomponent comprising at least one enzyme selected from the group consisting of hydrolases, subclass EC 3.8, and lyases, subclass EC 4.5, for carrying out a dehalogenation of the compound, said biocomponent immobilized to a surface of the tip by means selected from the group consisting of (a) entrapment within a hydrogel;
(b) entrapment within a polymeric network;
(c) (micro)encapsulation;
(d) covalent-bonding;
and (e) adsorption;
a treatment of said biocomponent for maintaining a period of enzymatic efficacy; and
said biocomponent stabilized by means selected from the group consisting of crosslinking a surface of said immobilized biocomponent, crosslinking a polymer layer to said biocomponent, adding a gel-hardening agent to said biocomponent, adding a stabilizing agent to said biocomponent, and modifying a component of said immobilizing means. - View Dependent Claims (2, 3, 4, 5)
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6. A distal tip of a biosensor ion sensing transducer for use in detecting an halogenated analyte selected from the group consisting of s-triazine compounds, gamma-hexachlorocyclohexane, and DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane), in an environment, the tip comprising:
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the ion-sensing transducer is selected from the group consisting of a pH optode, a pH electrode, a field-effect transistor (FET), and an halide ion-selective electrode (ISE);
a biocomponent comprising a dehalogenase for carrying out a dehalogenation of the analyte, said biocomponent immobilized to a surface of the tip by means selected from the group consisting of (a) entrapment within a hydrogel;
(b) entrapment within a polymeric network;
(c) (micro)encapsulation;
(d) covalent-bonding; and
(e) adsorption;
a treatment of said biocomponent for maintaining a period of enzymatic efficacy; and
said biocomponent stabilized by means selected from the group consisting of crosslinking a surface of said immobilized biocomponent, crosslinking a polymer layer to said biocomponent, adding a gel-hardening agent to said biocomponent, adding a stabilizing agent to said biocomponent, and modifying a component of said immobilizing means. - View Dependent Claims (7, 8, 9)
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10. A distal tip of a biosensor ion sensing transducer for use in detecting an analyte comprising an halogenated organic compound in an environment, the tip comprising:
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the ion-sensing transducer is selected from the group consisting of a pH optode, a pH electrode, a field-effect transistor (FET), and an halide ion-selective electrode (USE);
a biocomponent comprising a dehalogenase for carrying out a dehalogenation of the analyte, said biocomponent immobilized to a surface of the tip by entrapment of said dehalogenase within a matrix, said matrix selected from the group consisting of hydrogels and polymers;
a treatment of said biocomponent selected from the group consisting of applying an inhibitor of protein synthesis, adding a protease inhibitor, freeze drying, and dry heating; and
a polymer layer crosslinked to said biocomponent. - View Dependent Claims (11)
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12. A method of producing a biosensor distal tip comprising an ion sensing transducer and a biocomponent for use in detecting an analyte comprising an halogenated organic compound in an environment, the method comprising the steps of:
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immobilizing the biocomponent comprising at least one enzyme selected from the group consisting of hydrolases, subclass EC 3.8, and lyases, subclass EC 4.5, for carrying out a dehalogenation of the compound, to a surface of the tip by means selected from the group consisting of (a) entrapping said at least one enzyme within a hydrogel secured to said surface;
(b) entrapping said at least one enzyme within a polymeric network secured to said surface;
(c) (micro)encapsulating said at least one enzyme;
(d) covalent-bonding a second component of the biocomponent to said surface;
(e) cross-linking said at least one enzyme to a support material secured to said surface;
and (f) adsorbing said at least one enzyme into said surface;
treating the biocomponent for maintaining a period of enzymatic efficacy; and
stabilizing the biocomponent by means selected from the group consisting of crosslinking a polymer layer to the biocomponent, adding a gel-hardening agent to the biocomponent, adding a stabilizing agent to the biocomponent, and modifying a component of said immobilizing means. - View Dependent Claims (13)
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14. A method of producing a biosensor distal tip comprising an ion sensing transducer and a biocomponent for use in detecting an halogenated analyte selected from the group consisting of 6/1/O2-22of s-triazine compounds, gamma-hexachlorocyclohexane, and DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) in an environment, the method comprising the steps of:
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immobilizing the biocomponent comprising a dehalogenase for carrying out a dehalogenation of the analyte, to a surface of the tip by means selected from the group consisting of (a) entrapping said dehalogenase within a hydrogel secured to said surface;
(b) entrapping said dehalogenase within a polymeric network secured to said surface;
(c) microencapsulating said dehalogenase;
(d) covalent-bonding a second component of the biocomponent to said surface;
(e) cross-linking said dehalogenase to a support material secured to said surface; and
(f) adsorbing said dehalogenase into said surface; and
stabilizing the biocomponent by means selected from the group consisting of crosslinking a polymer layer to the biocomponent, adding a gel-hardening agent to the biocomponent, adding a stabilizing agent to the biocomponent, and modifying a component of said immobilizing means.
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15. A method of producing a biosensor distal tip comprising an ion sensing transducer and a biocomponent for use in detecting an analyte comprising an halogenated organic compound, the method comprising the steps of:
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immobilizing the biocomponent comprising a dehalogenase for carrying out a dehalogenation of the compound, to a surface of the tip by entrapping said dehalogenase within a hydrogel secured to said surface;
the ion-sensing transducer having been selected from the group consisting of a pH optode, a pH electrode, a field-effect transistor (FET), and an halide ion-selective electrode (ISE);
treating the biocomponent by means selected from the group consisting of (a) applying an inhibitor of protein synthesis to the biocomponent, said protein synthesis inhibitor selected from the group of antibiotics consisting of chloramphenicol, aminoglycosides, tetracyclines, and macrolides, (b) adding a protease inhibitor to the biocomponent, (c) freeze drying the biocomponent, and dry heating the biocomponent; and
crosslinking a polymer layer to the biocomponent.
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Specification