Electrochemical biosensors and process for their preparation
First Claim
Patent Images
1. An electrochemical biosensor comprising:
- a) an electro-conducting material, in the form of powder or grains;
b) a chemical mediator;
c) a substance absorbing said chemical mediator; and
d) one solid binding maker, selected from the group consisting of;
linear or branched, saturated or unsaturated hydrocarbons, containing from 12 to 60 carbon atoms optionally substituted with at least a group selected from —
OH, —
SH, —
NH2, —
CO—
. —
CHO, —
SO3H, —
COOH, —
OR1, —
SR1, —
NR1R2, and —
COOR1, wherein R1 and R2 are independently hydrocarbon groups, C1-C30, optionally containing one or more heteroatoms;
esters of fatty acid or a mixture of fatty acids with glycerol; and
esters of a fatty acid or a mixture of fatty acids with cholesterol, said solid binding maker having a melting point comprised between 35°
C. and 90°
C. and said elements a)-d) constituting a composite transducer, and a biocatalyst which is a redox enzyme, and wherein said biocatalyst is either incorporated in bulk into said composite transducer or it is applied in the form of a layer onto the surface of said composite transducer, said electrochemical biosensor being optionally covered by a membrane, and wherein said electrochemical biosensor comprises, with reference to 100 parts by weight of the sum of the components (a)-(d);
a) from 30 to 60% by weight;
b) from 1 to 10% by weight;
c) from 1 to 20% by weight;
d) from 30 to 60% by weight.
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Abstract
The present invention concerns new electrochemical biosensors for the determination of analytes concentration in sample solutions or suspensions, based on composite transducers containing an electro-conducting material, in the form of powder or grains, a chemical mediator, optionally a substance capable of sorption of said chemical mediator, and a solid binding maker, which is a compound in solid state at room temperature; said biosensors are prepared by incorporating a biocatalyst into the bulk of said composite transducers or by applying a biocatalytic layer onto their surface.
24 Citations
24 Claims
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1. An electrochemical biosensor comprising:
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a) an electro-conducting material, in the form of powder or grains;
b) a chemical mediator;
c) a substance absorbing said chemical mediator; and
d) one solid binding maker, selected from the group consisting of;
linear or branched, saturated or unsaturated hydrocarbons, containing from 12 to 60 carbon atoms optionally substituted with at least a group selected from —
OH, —
SH, —
NH2, —
CO—
. —
CHO, —
SO3H, —
COOH, —
OR1, —
SR1, —
NR1R2, and —
COOR1, wherein R1 and R2 are independently hydrocarbon groups, C1-C30, optionally containing one or more heteroatoms;
esters of fatty acid or a mixture of fatty acids with glycerol; and
esters of a fatty acid or a mixture of fatty acids with cholesterol, said solid binding maker having a melting point comprised between 35°
C. and 90°
C. and said elements a)-d) constituting a composite transducer, and a biocatalyst which is a redox enzyme, and wherein said biocatalyst is either incorporated in bulk into said composite transducer or it is applied in the form of a layer onto the surface of said composite transducer, said electrochemical biosensor being optionally covered by a membrane, and wherein said electrochemical biosensor comprises, with reference to 100 parts by weight of the sum of the components (a)-(d);
a) from 30 to 60% by weight;
b) from 1 to 10% by weight;
c) from 1 to 20% by weight;
d) from 30 to 60% by weight. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 24)
1) mixing an electro-conducting material with a chemical mediator;
2) optionally mixing said mixture obtained in step (1) with a substance adsorbing said chemical mediator;
3) optionally mixing said mixture obtained in step (1) or (2) with a biocatalyst;
4) mixing said mixture obtained in step (1), (2) or (3) with a solid binding maker;
5) introducing said mixture obtained in step (4) in a suitable holder, thus obtaining a compact form;
6) when step (3) is not worked out, applying a biocatalytic layer onto the surface of said transducer obtained in step (5); and
7) optionally covering said biosensor obtained in step (5) or (6) with a suitable membrane.
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15. The process according to claim 14 wherein, in step (4), said solid binding maker is mixed with said mixture obtained in step (1), (2) or (3) according to one of the following procedures:
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mechanical mixing;
mixing in the presence of a solvent which is then evaporated; and
melting said solid binding maker and mixing said mixture obtained in step (1), (2), or (3) with said solid binding maker in said melted state.
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24. The electrochemical biosensor according to claim 12, wherein said co-factor is selected from the group consisting of NAD, NADH, NADP, NADPH, FAD, FMN and quinone.
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16. An electrochemical biosensor comprising:
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a) an electro-conducting material, in the form of powder or grains;
b) a chemical mediator;
c) a substance absorbing said chemical mediator; and
d) one solid binding maker, selected from the group consisting of;
hexadecanol, hexadecanone, tetradecylamine, eicosane, tetracosane, monostearoyl glycerol, lecithin, cholesteryl myristate, cholesteryl stearate and cholesteryl oleate, said solid binding maker having a melting point comprised between 35°
C. and 90°
C. and said elements a)-d) constituting a composite transducer, and a biocatalyst which is a redox enzyme, and wherein said biocatalyst is either incorporated in bulk into said composite transducer or it is applied in the form of a layer onto the surface of said composite transducer, said electrochemical biosensor being optionally covered by a membrane, and wherein said electrochemical biosensor comprises, with reference to 100 parts by weight of the sum of the components (a)-(d);
a) from 30 to 60% by weight;
b) from 1 to 10% by weight;
c) from 1 to 20% by weight;
d) from 30 to 60% by weight.
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17. An electrochemical biosensor comprising:
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a) an electro-conducting material, in the form of powder or grains;
b) a chemical mediator;
c) a substance absorbing said chemical mediator; and
d) one solid binding maker, selected from the group consisting of;
linear or branched, saturated or unsaturated hydrocarbons, containing from 12 to 60 carbon atoms optionally substituted with at least a group selected from —
OH, —
SH, —
NH2, —
CO—
. —
CHO, —
SO3H, —
COOH, —
OR1, —
SR1, —
NR1R2, and —
COOR1, wherein R1 and R2 are independently hydrocarbon groups, C1-C30, optionally containing one or more heteroatoms;
esters of fatty acid or a mixture of fatty acids with glycerol; and
esters of a fatty acid or a mixture of fatty acids with cholesterol, said solid binding maker having a melting point comprised between 35°
C. and 90°
C. and said elements a)-d) constituting a composite transducer, and a biocatalyst which is composed of cells selected from the group consisting of Gluconobacter oxidans, Escherichia coli, Aspergillus niger, Pseudomonas fluorescens, Trichosporon brassicae, Saccharomyces cerevisiae, Breviabacterium lactofermentum, Enterobacter agglomerans, Leuconostoc mesenteroides, Nocardia erythropolys and mixtures thereof, and wherein said biocatalyst is either incorporated in bulk into said composite transducer or it is applied in the form of a layer onto the surface of said composite transducer, said electrochemical biosensor being optionally covered by a membrane, and wherein said electrochemical biosensor comprises, with reference to 100 parts by weight of the sum of the components (a)-(d);
a) from 30 to 60% by weight;
b) from 1 to 10% by weight;
c) from 1 to 20% by weight;
d) from 30 to 60% by weight.
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18. An electrochemical biosensor comprising:
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an electro-conducting material, in the form of powder or grains;
a chemical mediator;
optionally, a substance adsorbing said chemical mediator;
one solid binding maker selected from the group consisting of hexadecanol, hexadecanone, tetradecylamine, eicosane, tetracosane, monostearoyl glycerol, lecithin, cholesteryl myristate, cholesteryl stearate, or cholesteryl oleate; and
a biocatalyst which is a redox enzyme, wherein said biocatalyst is either incorporated in bulk into a composite transducer or it is applied in the form of a layer onto the surface of said composite transducer, said electrochemical biosensor being optionally covered by a membrane. - View Dependent Claims (19, 20)
1) mixing an electro-conducting material with a chemical mediator;
2) optionally mixing said mixture obtained in step (1) with a substance adsorbing said chemical mediator;
3) optionally mixing said mixture obtained in step (1) or (2) with a biocatalyst;
4) mixing said mixture obtained in step (1), (2) or (3) with a solid binding maker;
5) introducing said mixture obtained in step (4) in a suitable holder, thus obtaining a compact form;
6) when step (3) is not worked out, applying a biocatalytic layer onto the surface of said transducer obtained in step (5); and
7) optionally covering said biosensor obtained in step (5) or (6) with a suitable membrane.
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20. The process according to claim 19 wherein, in step (4), said solid binding maker is mixed with said mixture obtained in step (1), (2) or (3) according to one of the following procedures:
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mechanical mixing;
mixing in the presence of a solvent which is then evaporated; and
melting said solid binding maker and mixing said mixture obtained in step (1), (2), or (3) with said solid binding maker in said melted state.
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21. An electrochemical biosensor comprising:
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a) an electro-conducting material, in the form of powder or grains;
b) a chemical mediator;
c) optionally, a substance adsorbing said chemical mediator;
d) one solid binding maker, selected from the group of;
linear or branched, saturated or unsaturated hydrocarbons, containing from 12 to 60 carbon atoms optionally substituted with at least a group selected from —
OH, —
SH, —
NH2, —
CO—
, —
CHO, —
SO3H, —
COOH, —
OR1, —
SR1, —
NR1R2 and —
COOR1, wherein R1 and R2 are independently hydrocarbon groups C1-C30, optionally containing one or more heteroatoms;
esters of a fatty acid or a mixture of fatty acids with glycerol; and
esters of a fatty acid or a mixture of fatty acids with cholesterol, said solid binding maker having a melting point comprised between 35°
C. and 90°
C. and said elements a)-d) constituting a composite transducer, and a biocatalyst composed of cells selected from the group consisting of Gluconobacter oxidans, Escherichia coli, Aspergillus niger, Pseudomonas fluorescens, Trichosporon brassicae, Saccharomyces cerevisiae, Breviacterium lactofermentum, Enterobacter agglomerans, Leuconostoc mesenteroides, Nocardia erythropolys and mixtures thereof.- View Dependent Claims (22, 23)
1) mixing an electro-conducting material with a chemical mediator;
2) optionally mixing said mixture obtained in step (1) with a substance adsorbing said chemical mediator;
3) optionally mixing said mixture obtained in step (1) or (2) with a biocatalyst;
4) mixing said mixture obtained in step (1), (2) or (3) with a solid binding maker;
5) introducing said mixture obtained in step (4) in a suitable holder, thus obtaining a compact form;
6) when step (3) is not worked out, applying a biocatalytic layer onto the surface of said transducer obtained in step (5); and
7) optionally covering said biosensor obtained in step (5) or (6) with a suitable membrane.
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23. The process according to claim 22 wherein, in step (4), said solid binding maker is mixed with said mixture obtained in step (1), (2) or (3) according to one of the following procedures:
-
mechanical mixing;
mixing in the presence of a solvent which is then evaporated; and
melting said solid binding maker and mixing said mixture obtained in step (1), (2), or (3) with said solid binding maker in said melted state.
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Specification