WO2001040452A2 - Biologically active material - Google Patents

Biologically active material Download PDF

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Publication number
WO2001040452A2
WO2001040452A2 PCT/EE2000/000004 EE0000004W WO0140452A2 WO 2001040452 A2 WO2001040452 A2 WO 2001040452A2 EE 0000004 W EE0000004 W EE 0000004W WO 0140452 A2 WO0140452 A2 WO 0140452A2
Authority
WO
WIPO (PCT)
Prior art keywords
biologically active
active material
bioactive component
amount
bioactive
Prior art date
Application number
PCT/EE2000/000004
Other languages
French (fr)
Other versions
WO2001040452A3 (en
WO2001040452B1 (en
Inventor
Toonika Rinken
Jaak Järv
Ago Rinken
Toomas Tenno
Original Assignee
University Of Tartu
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Tartu filed Critical University Of Tartu
Priority to AU15135/01A priority Critical patent/AU1513501A/en
Publication of WO2001040452A2 publication Critical patent/WO2001040452A2/en
Publication of WO2001040452A3 publication Critical patent/WO2001040452A3/en
Publication of WO2001040452B1 publication Critical patent/WO2001040452B1/en

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof

Definitions

  • the present invention belongs to the field of chemical analysis and chemical catalysis and is meant for application in medical and environmental analyses, biotechnological industry, scientific research and other fields .
  • the biologically active material consists of a bioactive component e.g. microorganisms, enzymes, antibodies, etc. and an insoluble carrier of the bioactive component e.g. a mesh, a film and analogous.
  • a bioactive component e.g. microorganisms, enzymes, antibodies, etc.
  • an insoluble carrier of the bioactive component e.g. a mesh, a film and analogous.
  • the biologically active material (the insoluble carrier with the attached bioactive component) is used for the preparation of biosensors for the determination of various compounds (US patent No 5637201, G01N 027/26, B.Raguse, B.-A. Georgia, V.-I. Braach-Ma svytis , R.J. Pace, 1995), but also as catalysts to speed up different processes in industry and other fields of application
  • the insoluble carrier to which the bioactive component is attached is in the form of films (US patent No 4871440, G 01 N 027/54, Y.Nagata, H.Fujimura, 1989) or various porous materials (membranes) (US patent No 4418148, C12N 011/12, B.Oberhardt 1981) .
  • the disadvantage of prior art is the invariability of the amount of the bioactive component attached to one unit of the insoluble carrier due to the two-dimensional plane fixation of the carrier.
  • the biologically active material where the insoluble carrier for the attachment of bioactive material has the form of a mesh (US patent No 4929330, G01N 027/30 O.Tatsuhiko, T.Hiroshi ja O.Mitsunari, 1990) .
  • the planar fixed form of the carrier does not allow the variation of the amount of the bioactive component corresponding to one unit of the biologically active material .
  • the specificity and selectivity of the biologically active material are determined by the characteristics of the bioactive component.
  • the amount of bioactive component determines the efficiency of the biologically active material.
  • the bioactive component and its amount are not easily variable using the planar fixed carrier.
  • the present invention proposes a biologically active material consisting of a bioactive component and its carrier, which has the form of a thread.
  • Such a form of the carrier enables us to measure the amount of the carrier and the amount of the bioactive component in the units of length and to cut pieces of a biologically active material in which the amount of the bioactive component is measurable in the units of length.
  • the bioactive component in the biologically active material and its amount are variable. Additionally, the biologically active material can contain several different bioactive components.
  • the form of the thread of the carrier, consistently containing a bioactive component enables us to measure the amount of the bioactive component in the units of length, e.g. with a ruler.
  • Thread-shaped biologically active material was used for the construction of glucose biosensors and the determination of glucose concentration in solutions.
  • the glucose biosensor consists of the amperometric oxygen sensor with the cylindrical cathode and the thread-shaped biologically active material consisting glucose oxidase.
  • Glucose oxidase is the enzyme which catalyses the oxidation reaction of glucose by oxygen.
  • the cylindrical cathode of the amperometric oxygen sensor had the area of 5,65 cm 2 and was covered with 60 ⁇ m thick polyethylene.
  • Glucose oxidase was attached to the nylon-6.6 thread with the help of glutaraldehyde . From this biologically active material (consisting of glucose oxidase and nylon carrier) the 80 cm long fragment corresponding to 0,9 ⁇ g soluble glucose oxidase was cut .
  • the biologically active material i.e. glucose oxidase containing nylon thread, was wound spirally around the outer surface of polyethylene membrane, covering the cathode of the amperometric oxygen sensor so, that it covered 50 % of the cathode membrane surface.
  • Threads with glucose oxidase of different length were used for the determination of glucose concentration. So the amount of glucose oxidase was variable and could be measured in the units of length.
  • the determination range of glucose concentration could be selected from 0-1 mmol/1 (with uncertainty of experiment ⁇ 0.02mmol/l) up to 0- 40 mmol/1 (with uncertainty of experiment ⁇ 1.5 mmol/1) .
  • the present invention was also used for the determination of other compounds and their concentrations e.g. the uric acid and its salts, catechol, phenol) .
  • the threads containing other bioactive components uricase, tyrosinase were used in biosensors .
  • the bioactive component containing thread To place and replace the biologically active material (the bioactive component containing thread) easily.

Abstract

Biologically active material is consisting of a bioactive component and its thread-shaped carrier. The thread-shaped carrier enables us to measure the amount of the bioactive component in units of length and cut pieces of biologically active material, the bioactive material content of which is measured in units of length. In the biologically active material, the bioactive component and its amount are variable and the biologically active material can contain several different bioactive components.

Description

BIOLOGICALLY ACTIVE MATERIAL
TECHNICAL FIELD
The present invention belongs to the field of chemical analysis and chemical catalysis and is meant for application in medical and environmental analyses, biotechnological industry, scientific research and other fields .
BACKGROUND ART
The biologically active material consists of a bioactive component e.g. microorganisms, enzymes, antibodies, etc. and an insoluble carrier of the bioactive component e.g. a mesh, a film and analogous.
The biologically active material (the insoluble carrier with the attached bioactive component) is used for the preparation of biosensors for the determination of various compounds (US patent No 5637201, G01N 027/26, B.Raguse, B.-A. Cornell, V.-I. Braach-Ma svytis , R.J. Pace, 1995), but also as catalysts to speed up different processes in industry and other fields of application
(Pialis, Hamann et al . L-DOPA Production from Tyrosinase
Immobilized on Nylon 6,6. Biotechnology and
Bioengineering, Vol. 51, p. 141-147, 1996).
Usually the insoluble carrier to which the bioactive component is attached is in the form of films (US patent No 4871440, G 01 N 027/54, Y.Nagata, H.Fujimura, 1989) or various porous materials (membranes) (US patent No 4418148, C12N 011/12, B.Oberhardt 1981) . The disadvantage of prior art is the invariability of the amount of the bioactive component attached to one unit of the insoluble carrier due to the two-dimensional plane fixation of the carrier. To the present invention most similar biologically active material from among the known biologically active materials is the biologically active material, where the insoluble carrier for the attachment of bioactive material has the form of a mesh (US patent No 4929330, G01N 027/30 O.Tatsuhiko, T.Hiroshi ja O.Mitsunari, 1990) . However, the planar fixed form of the carrier does not allow the variation of the amount of the bioactive component corresponding to one unit of the biologically active material .
The specificity and selectivity of the biologically active material are determined by the characteristics of the bioactive component. The amount of bioactive component determines the efficiency of the biologically active material. The bioactive component and its amount are not easily variable using the planar fixed carrier.
DISCLOSURE OF THE INVENTION
The present invention proposes a biologically active material consisting of a bioactive component and its carrier, which has the form of a thread.
Such a form of the carrier enables us to measure the amount of the carrier and the amount of the bioactive component in the units of length and to cut pieces of a biologically active material in which the amount of the bioactive component is measurable in the units of length.
The bioactive component in the biologically active material and its amount are variable. Additionally, the biologically active material can contain several different bioactive components.
The form of the thread of the carrier, consistently containing a bioactive component enables us to measure the amount of the bioactive component in the units of length, e.g. with a ruler.
To get a definite amount of the bioactive component, a piece of biologically active material with a definite length has to be cut.
DESCRIPTION OF EMBODIMENT
Thread-shaped biologically active material was used for the construction of glucose biosensors and the determination of glucose concentration in solutions. The glucose biosensor consists of the amperometric oxygen sensor with the cylindrical cathode and the thread-shaped biologically active material consisting glucose oxidase. Glucose oxidase is the enzyme which catalyses the oxidation reaction of glucose by oxygen.
The cylindrical cathode of the amperometric oxygen sensor had the area of 5,65 cm2 and was covered with 60 μm thick polyethylene.
Glucose oxidase was attached to the nylon-6.6 thread with the help of glutaraldehyde . From this biologically active material (consisting of glucose oxidase and nylon carrier) the 80 cm long fragment corresponding to 0,9 μg soluble glucose oxidase was cut . The biologically active material i.e. glucose oxidase containing nylon thread, was wound spirally around the outer surface of polyethylene membrane, covering the cathode of the amperometric oxygen sensor so, that it covered 50 % of the cathode membrane surface.
All the measurements were carried out in thermostated (25° C) glass cells in buffered solutions (pH=5.60) under homogeneous stirring. These conditions were optimal to achieve the maximum activity and stability of glucose oxidase. The biosensor output data were registered and processed automatically with a computer. It was possible to determine the glucose concentration in the range 0-2 mmol/1 (± 0.05mmol/l) with the above-described biosensor.
Threads with glucose oxidase of different length (60, 100, 120 cm) were used for the determination of glucose concentration. So the amount of glucose oxidase was variable and could be measured in the units of length. The determination range of glucose concentration could be selected from 0-1 mmol/1 (with uncertainty of experiment ± 0.02mmol/l) up to 0- 40 mmol/1 (with uncertainty of experiment ± 1.5 mmol/1) .
To broaden the range of determination, along with a glucose oxidase containing thread, threads without this bioactive component was used in biosensors.
The present invention was also used for the determination of other compounds and their concentrations e.g. the uric acid and its salts, catechol, phenol) . For these processes of determination the threads containing other bioactive components (uricase, tyrosinase) were used in biosensors .
The use of the proposed biologically active material enables:
To measure the amount of a bioactive component in the units of length in the case of the homogeneous distribution of the bioactive component throughout the thread; - To cut pieces of biologically active material which contain the measurable amount of bioactive component in the units of length;
To place and replace the biologically active material (the bioactive component containing thread) easily.

Claims

1. Biologically active material consisting of a bioactive component and its carrier c h a r a c t e r i z e d in this that the carrier has the shape of a thread, allowing to measure the amount of the biologically active material and the amount of the bioactive component in it in the units of length and to cut pieces of biologically active material, which contain a stated amount of bioactive component measurable in the units of length.
2. Biologically active material according to claim 1, c h a r a c t e r i z e d in this that the bioactive component and its amount on the thread-shaped carrier are variable .
3. Biologically active material according to claim 1, c h a r a c t e r i z e d in this that it contains several different bioactive components.
PCT/EE2000/000004 1999-11-29 2000-11-27 Biologically active material WO2001040452A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU15135/01A AU1513501A (en) 1999-11-29 2000-11-27 Biologically active material

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EEP199900442A EE9900442A (en) 1999-11-29 1999-11-29 Biologically active material
EEP199900442 1999-11-29

Publications (3)

Publication Number Publication Date
WO2001040452A2 true WO2001040452A2 (en) 2001-06-07
WO2001040452A3 WO2001040452A3 (en) 2001-11-08
WO2001040452B1 WO2001040452B1 (en) 2002-07-25

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PCT/EE2000/000004 WO2001040452A2 (en) 1999-11-29 2000-11-27 Biologically active material

Country Status (3)

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AU (1) AU1513501A (en)
EE (2) EE04250B1 (en)
WO (1) WO2001040452A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2529217B1 (en) 2010-01-29 2013-10-23 Tartu Ülikool (University Of Tartu) On-line system, method of its calibration and simultaneous detection of antibiotic residues and their concentration in milk

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0183184A1 (en) * 1984-11-30 1986-06-04 Ppg Industries, Inc. Method of cultivating cellular biomaterial and glass fibers with cellular biomaterial or biomaterial
EP0205232A1 (en) * 1985-04-08 1986-12-17 Kelsius, Inc. Amplification of signals from optical fibers
EP0328256A1 (en) * 1988-01-21 1989-08-16 Owens-Corning Fiberglas Corporation Glass fibers coated with agarose for use as column packing or chromatographic media for bioseparations
US5308889A (en) * 1988-11-21 1994-05-03 Collagen Corporation Dehydrated collagen-polymer strings
US5837196A (en) * 1996-01-26 1998-11-17 The Regents Of The University Of California High density array fabrication and readout method for a fiber optic biosensor

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0183184A1 (en) * 1984-11-30 1986-06-04 Ppg Industries, Inc. Method of cultivating cellular biomaterial and glass fibers with cellular biomaterial or biomaterial
EP0205232A1 (en) * 1985-04-08 1986-12-17 Kelsius, Inc. Amplification of signals from optical fibers
EP0328256A1 (en) * 1988-01-21 1989-08-16 Owens-Corning Fiberglas Corporation Glass fibers coated with agarose for use as column packing or chromatographic media for bioseparations
US5308889A (en) * 1988-11-21 1994-05-03 Collagen Corporation Dehydrated collagen-polymer strings
US5837196A (en) * 1996-01-26 1998-11-17 The Regents Of The University Of California High density array fabrication and readout method for a fiber optic biosensor

Also Published As

Publication number Publication date
EE9900442A (en) 2001-08-15
AU1513501A (en) 2001-06-12
EE04250B1 (en) 2004-02-16
WO2001040452A3 (en) 2001-11-08
WO2001040452B1 (en) 2002-07-25
EE200200674A (en) 2003-04-15

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