EP0228259A2
Expired / Lapsed
EP
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Enzyme immobilized membrane for a semiconductor sensor and method for producing same
Assignee: Seitaikinouriyou Kagakuhin Sinseizogijutsu Kenkyu Kumiai
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Filing: 1986-12-18
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Granted: —
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Expires: 2006-12-18
Abstract
Abstract An enzyme immobilized membrane and a method for producing the same are disclosed in which the portion of an enzyme membrane having photo crosslinking, even if containing an excessive amount of an enzyme, does not peel off from a sensitive portion of an enzyme sensor. Elution of the enzyme from the enzyme membrane can be effectively prevented, and an enzyme immobilized membrane having a limited area and high mechanical strength can be produced with good reproduceability. An aqueous solution, composed of a water soluble photosensitive resin including polyvinyl pyrrolidone (109) and a cyclopentanone sodium salt (110) and an enzyme (102), is coated on a base (101) and dried so as to form an enzyme membrane. Then, light of an appropriate wavelength such as ultraviolet light is irradiated on the enzyme membrane to provide photo crosslinking between the polyvinyl pyrrolidone, the cyclopentanone sodium salt, and the enzyme. Such enzyme immobilized membranes can be directly formed on the ion-sensitive portions of pH-ISFETs so as to form a semiconductor enzyme sensor.
Inventors
- 👤 Yoshio Chuo Kenkyusho Of Mitsubishi Hanazato
- 👤 Satoru C/O Chuo Kenkyusho Misubishi Shiono
- 👤 Mamiko§c/o Chuo Kenkyusho§Mitsubishi Nakako
- 👤 Satoshi C/O Chuo Kenkyusho Misubishi Yamada
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-02 15:51
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/001; C12Q1/002; C12N; C12N11/00; C12N11/02; C12N11/08; C12N11/082; C12N11/089; C12N11/091; C12N11/098; Y; Y10; Y10S; Y10S435/00; Y10S435/817
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
36 words
An enzyme immobilized membrane, characterized by comprising: a water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2 ' -sulfobenzal)cyclopentanone sodium salt; and an enzyme.
Claim 2
Dependent Claim
36 words
An enzyme immobilized membrane, characterized by comprising: a water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2 ' -sulfobenzal)cyclopentanone sodium salt; and an enzyme.
Claim 3
Dependent Claim
23 words
An enzyme immobilized membrane as claimed in claim 1, characterized in that the said enzyme is selected from glucose oxidase, ureaze and lipase.
Claim 4
Dependent Claim
28 words
An enzyme immobilized membrane as claimed in claim 1 or 2, characterized by further comprising: glutaraldehyde; and a material adapted to cooperate with glutaraldehyde to form chemical crosslinking.
Claim 5
Dependent Claim
36 words
An enzyme immobilized membrane as claimed in claim 3, characterized in that the said material adapted to cooperate with glutaraldehyde to form chemical crosslinking is selected from bovine serum albumin, polyamino acid, and polyamino acid copolymer.
Claim 6
Dependent Claim
18 words
An enzyme immobilized membrane as claimed in claim 4, characterized in that the said polyamino acid is poly-L-lysine.
Claim 7
Dependent Claim
24 words
An enzyme immobilized membrane as claimed in claim 4, characterized in that the said polyamino acid copolymer is a copolymer of poly-L-lysine and poly-L-thyrosin.
Claim 8
Dependent Claim
75 words
An enzyme immobilized membrane as claimed in any of claims 1 to 6, characterized in that the said polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands comprises 2-20 weight %; and said 2,5-bis(4'-azide-2'-sulfobenzal) cyclopentanone sodium salt comprises 0.5 - 1.5 weight %; and said enzyme comprises glucose oxidase of 5.0 - 7.5 weight %; said enzyme immobilized membrane further comprising bovine serum albumin of 5 - 10 weight %.
Claim 9
Independent Claim
94 words
A method for producing an enzyme immobilized membrane, characterized by comprising the steps of: coating on a base an aqueous solution which is composed of a water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2'-sulfobenzal)cyclopentanone sodium salt, and an enzyme; drying the aqueous solution thus coated on the said base so as to form an enzyme immobilized membrane; and irradiating ultraviolet light on the said enzyme immobilized membrane to provide photo-crosslinking between the polyvinyl pyrrolidone, the cyclopentanone sodium salt, and the enzyme.
Claim 10
Independent Claim
94 words
A method for producing an enzyme immobilized membrane, characterized by comprising the steps of: coating on a base an aqueous solution which is composed of a water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2'-sulfobenzal)cyclopentanone sodium salt, and an enzyme; drying the aqueous solution thus coated on the said base so as to form an enzyme immobilized membrane; and irradiating ultraviolet light on the said enzyme immobilized membrane to provide photo-crosslinking between the polyvinyl pyrrolidone, the cyclopentanone sodium salt, and the enzyme.
Claim 11
Dependent Claim
21 words
A method as claimed in claim 8, characterized in that the said enzyme is selected from glucose oxidase, ureaze and lipase.
Claim 12
Dependent Claim
31 words
A method as claimed in claim 8 or 9, characterized in that the said aqueous solution further comprises: glutaraldehyde: and a material adapted to cooperate with glutaraldehyde to form chemical crosslinking.
Claim 13
Dependent Claim
34 words
A method as claimed in claim 10, characterized in that the said material adapted to cooperate with glutaraldehyde to form chemical crosslinking is selected from bovine serum albumin, polyamino acid, and polyamino acid copolymer.
Claim 14
Dependent Claim
16 words
A method as claimed in claim 11, characterized in that the said polyamino acid is poly-L-lysine.
Claim 15
Dependent Claim
22 words
A method as claimed in claim 11, characterized in that the said polyamino acid copolymer is a copolymer of poly-L-lysine and poly-L-thyrosin.
Claim 16
Independent Claim
71 words
A method as claimed in any of claims 8 to 13, characterized in that said polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands comprises 2 - 20 weight %; and said 2,5-bis(4'-azide-2'-sulfobenzal)cyclopentanone sodium salt comprises 0.5 - 1.5 weight%; and said enzyme comprises glucose oxidase of 5.0 - 7.5 weight %: said aqueous solution further comprising bovine serum albumin of 5 - 10 weight %.
Claim 17
Independent Claim
37 words
A method as claimed in any of claims 8 to 14, characterized by further comprising dipping said enzyme immobilized membrane irradiated by said ultraviolet light into an aqueous solution of glutaraldehyde so as to form chemical crosslinking.
Claim 18
Dependent Claim
21 words
A method as claimed in claim 15, characterized by further comprising washing away the glucose-oxidase immobilized membrane by a buffer solution.
Claim 19
Dependent Claim
46 words
A method as claimed in claim 16, characterized by further comprising: dipping said glucose-oxidase immobilized membrane once washed away by said buffer solution into an aqueous solution of glycine; and washing away said glucose-oxidase immobilised membrane by a buffer solution for removal of the residual glutaraldehyde.
Claim 20
Dependent Claim
63 words
A semiconductor enzyme sensor, characterized by comprising: a first pH-ISFET having a first enzyme immobilized membrane connected thereto, the said first membrane being composed of a first water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2 1- sulfobenzal)cyclopentanone sodium salt, and a first enzyme: a second pH-ISFET: and a reference electrode.
Claim 21
Dependent Claim
63 words
A semiconductor enzyme sensor, characterized by comprising: a first pH-ISFET having a first enzyme immobilized membrane connected thereto, the said first membrane being composed of a first water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2 1- sulfobenzal)cyclopentanone sodium salt, and a first enzyme: a second pH-ISFET: and a reference electrode.
Claim 22
Dependent Claim
60 words
A semiconductor enzyme sensor as claimed in claim 18, characterized by further comprising a third pH-ISFET having a second enzyme immobilized membrane connected thereto, the said second membrane being composed of a second water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2'-sulfobenzal)cyclopentanone sodium salt, and a second enzyme.
Claim 23
Dependent Claim
24 words
A semiconductor enzyme sensor as claimed in claim 19, characterized in that the said second enzyme is selected from glucose oxidase, ureaze and lipase.
Claim 24
Dependent Claim
28 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 20, characterized in that the said first enzyme is selected from glucose oxidase, ureaze and lipase.
Claim 25
Dependent Claim
30 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 21, characterized by further comprising: glutaraldehyde: and a material adapted to cooperate with glutaraldehyde to form chemical crosslinking.
Claim 26
Dependent Claim
36 words
A semiconductor enzyme sensor as claimed in claim 22, characterized in that the said material adapted to cooperate with glutaraldehyde to form chemical crosslinking is selected from bovine serum albumin, polyamino acid, and polyamino acid copolymer.
Claim 27
Dependent Claim
18 words
A semiconductor enzyme sensor as claimed in claim 23, characterized in that the said polyamino acid is poly-L-lysine.
Claim 28
Dependent Claim
24 words
A semiconductor enzyme sensor as claimed in claim 23, characterized in that the said polyamino acid copolymer is a copolymer of poly-L-lysine and poly-L-thyrosin.
Claim 29
Dependent Claim
77 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 25, characterized in that the said polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands comprises 2 - 20 weight %; and said 2,5-bis(4'-azide-2'-sulfobenzal) cyclopentanone sodium salt comprises 0.5 - 1.5 weight %; and said enzyme comprises glucose oxidase of 5.0 - 7.5 weight %; said enzyme immobilized membrane further comprising bovine serum albumin of 5 - 10 weight %.
Claim 30
Dependent Claim
30 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 26, characterized in that the said first enzyme immobilized membrane is directly coated and hardened on said pH-ISFET.
Claim 31
Dependent Claim
41 words
A semiconductor enzyme sensor as claimed in claim 19, or as claimed in any of claims 20 to 27 when appendant to claim 19, characterized in that the said second enzyme immobilized membrane is directly coated and hardened on said pH-ISFET.
Claim 32
Dependent Claim
77 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 28, characterized in that the said first enzyme immobilized membrane is formed by coating on a pH-ISFET a first aqueous solution containing said first water soluble photosensitive resin, glucose oxidase, bovine serum albumin, and hardening said first aqueous solution thus coated by irradiation of ultraviolet light, said first enzyme immobilized membrane being dipped into a glutaraldehyde solution to form chemical crosslinking for improved mechanical strength.
Claim 33
Dependent Claim
88 words
A semiconductor enzyme sensor as claimed in claim 19, or as claimed in any of claims 20 to 29 when appendant to claim 19, characterized in that the said second enzyme immobilized membrane is formed by coating on a pH-ISFET a second aqueous solution containing said second water soluble photosensitive resin, glucose oxidase, bovine serum albumin, and hardening said second aqueous solution thus coated by irradiation of ultraviolet light, said second enzyme immobilized membrane being dipped into a glutaraldehyde solution to form chemical crosslinking for improved mechanical strength.
Claim 34
Dependent Claim
47 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 30, characterized in that the said first pH-ISFET has an ion-sensitive surface, and said first enzyme immobilized membrane is formed through patterning on the ion-sensitive surface of said first pH-ISFET by m anε of photolithography.
Claim 35
Dependent Claim
57 words
A semiconductor enzyme sensor as claimed in claim 19, or as claimed in any of claims 20 to 31 when appendant to claim 19. characterized in that the said third pH-ISFET has an ion-sensitive surface, and said second enzyme immobilized membrane is formed through patterning on the ion-sensitive surface of said third pH-ISFET by means of photolithography.
Claim 36
Dependent Claim
66 words
A semiconductor enzyme sensor as claimed in any of claims 18 to 32, characterized by further comprising further one or more pH-ISFETs each having a further enzyme immobilized membrane connected thereto, said further membrane being composed of a further water soluble photosensitive resin including polyvinyl pyrrolidone having a molecular weight of tens of thousands to hundreds of thousands and 2,5-bis(4'-azide-2'-sulfobenzal)cyclopentanone sodium salt, and a further enzyme.
References Cited (Backward Citations)
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