Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes

Timo Mäkelä, Martin Albers

Research output: Book/ReportReportProfessional

Abstract

This project report deals with the construction of amperometric biosensors for the detection of choline and lactate, using Choline Oxidase and Lactate Oxidase as the selective element. The enzymes were immobilised in a variety of microporous membrane supports using different immobilisation protocols. Microporous membranes for immobilisation of biologically active materials are available commercially from Millipore (USA) as "Immobilon" and Pall (UK) as Pall "Biodyne". Also "track-etch" membranes (Cyclopore SA, Belgium) were used in the comparison as a promising new support. The electrochemical sensor response of the membranes was compared with a spectrophotometric determination of enzyme activity for the various immobilisation methods used. The best membrane for immobilisation appeared to be the track-etch membrane, which gave a fast response and a good reproducibility. The problem with these membranes, however, in the case of ChOx immobilisation was the saturation of the choline response in the measurement range. Generally, the Immobilon and Pall membranes have a larger binding capacity, but give lower reproducibility as the track-etch membranes. A more designed approach to effective immobilisation is in principle possible when the crystal structure of an enzyme is available. The exact structure of lactate oxidase, however, is still unknown. In this study the immobilisation of enzymes has been studied merely in an empirical fashion. The large differences observed with various methods and membranes were quite striking and highlight the importance of choosing the right materials and methods for immobilisation of enzymes.
Original languageEnglish
Place of PublicationEspoo
PublisherVTT Technical Research Centre of Finland
Number of pages44
ISBN (Print)951-38-4970-8
Publication statusPublished - 1996
MoE publication typeNot Eligible

Publication series

NameVTT Tiedotteita - Meddelanden - Research Notes
PublisherVTT
No.1775
ISSN (Print)1235-0605
ISSN (Electronic)1455-0865

Fingerprint

lactate 2-monooxygenase
choline oxidase
Choline
Biosensors
Lactic Acid
Membranes
Enzymes
Electrochemical sensors

Keywords

  • bioinstrumentation
  • detectors
  • electrical measurement
  • measurement
  • cholines
  • lactates
  • immobilization
  • microporosity
  • membranes
  • enzymes

Cite this

Mäkelä, T., & Albers, M. (1996). Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes. Espoo: VTT Technical Research Centre of Finland. VTT Tiedotteita - Meddelanden - Research Notes, No. 1775
Mäkelä, Timo ; Albers, Martin. / Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes. Espoo : VTT Technical Research Centre of Finland, 1996. 44 p. (VTT Tiedotteita - Meddelanden - Research Notes; No. 1775).
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Mäkelä, T & Albers, M 1996, Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes. VTT Tiedotteita - Meddelanden - Research Notes, no. 1775, VTT Technical Research Centre of Finland, Espoo.

Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes. / Mäkelä, Timo; Albers, Martin.

Espoo : VTT Technical Research Centre of Finland, 1996. 44 p. (VTT Tiedotteita - Meddelanden - Research Notes; No. 1775).

Research output: Book/ReportReportProfessional

TY - BOOK

T1 - Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes

AU - Mäkelä, Timo

AU - Albers, Martin

PY - 1996

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N2 - This project report deals with the construction of amperometric biosensors for the detection of choline and lactate, using Choline Oxidase and Lactate Oxidase as the selective element. The enzymes were immobilised in a variety of microporous membrane supports using different immobilisation protocols. Microporous membranes for immobilisation of biologically active materials are available commercially from Millipore (USA) as "Immobilon" and Pall (UK) as Pall "Biodyne". Also "track-etch" membranes (Cyclopore SA, Belgium) were used in the comparison as a promising new support. The electrochemical sensor response of the membranes was compared with a spectrophotometric determination of enzyme activity for the various immobilisation methods used. The best membrane for immobilisation appeared to be the track-etch membrane, which gave a fast response and a good reproducibility. The problem with these membranes, however, in the case of ChOx immobilisation was the saturation of the choline response in the measurement range. Generally, the Immobilon and Pall membranes have a larger binding capacity, but give lower reproducibility as the track-etch membranes. A more designed approach to effective immobilisation is in principle possible when the crystal structure of an enzyme is available. The exact structure of lactate oxidase, however, is still unknown. In this study the immobilisation of enzymes has been studied merely in an empirical fashion. The large differences observed with various methods and membranes were quite striking and highlight the importance of choosing the right materials and methods for immobilisation of enzymes.

AB - This project report deals with the construction of amperometric biosensors for the detection of choline and lactate, using Choline Oxidase and Lactate Oxidase as the selective element. The enzymes were immobilised in a variety of microporous membrane supports using different immobilisation protocols. Microporous membranes for immobilisation of biologically active materials are available commercially from Millipore (USA) as "Immobilon" and Pall (UK) as Pall "Biodyne". Also "track-etch" membranes (Cyclopore SA, Belgium) were used in the comparison as a promising new support. The electrochemical sensor response of the membranes was compared with a spectrophotometric determination of enzyme activity for the various immobilisation methods used. The best membrane for immobilisation appeared to be the track-etch membrane, which gave a fast response and a good reproducibility. The problem with these membranes, however, in the case of ChOx immobilisation was the saturation of the choline response in the measurement range. Generally, the Immobilon and Pall membranes have a larger binding capacity, but give lower reproducibility as the track-etch membranes. A more designed approach to effective immobilisation is in principle possible when the crystal structure of an enzyme is available. The exact structure of lactate oxidase, however, is still unknown. In this study the immobilisation of enzymes has been studied merely in an empirical fashion. The large differences observed with various methods and membranes were quite striking and highlight the importance of choosing the right materials and methods for immobilisation of enzymes.

KW - bioinstrumentation

KW - detectors

KW - electrical measurement

KW - measurement

KW - cholines

KW - lactates

KW - immobilization

KW - microporosity

KW - membranes

KW - enzymes

M3 - Report

SN - 951-38-4970-8

T3 - VTT Tiedotteita - Meddelanden - Research Notes

BT - Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -

Mäkelä T, Albers M. Amperometric biosensors for detection of choline and lactate by Choline Oxidase and Lactate Oxidase immobilisation in microporous membranes. Espoo: VTT Technical Research Centre of Finland, 1996. 44 p. (VTT Tiedotteita - Meddelanden - Research Notes; No. 1775).