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Proceedings Paper

Flow injection analysis with bioluminescence-based fiber-optic biosensors
Author(s): Loic J. Blum; Sabine Gautier; Pierre R. Coulet
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Paper Abstract

Fiber optic biosensors based on the firefly and the bacterial bioluminescence reactions have been constructed and incorporated in a specially designed flow-cell for the sensitive determination of ATP and NADH, respectively. The bioluminescence enzymes were immobilized on preactivated polyamide membranes which were placed in close contact with the surface on one end of a glass-fiber bundle, the other end being connected to the photomultiplier tube of a luminometer. When using the continuous-flow device with the firefly luciferase or the bacterial system immobilized separately on different membranes, the detection limit for ATP and NADH were 0.25 and 2 pmol, respectively. The versatility of the fiber optic probe has been improved by co-immobilizing the bacterial bioluminescent system and the firefly luciferase on the same support enabling the use of a single sensor for the selective, specific, and alternate determination of these two analytes. Compatible reaction conditions preserving the activity of each co-immobilized enzyme without impairing its stability were found. The selection of the appropriate reaction medium was done using a four port valve. Alternate quantification of ATP and NADH could then be performed in the linear ranges 0.25 pmol - 3 nmol and 5 pmol - 1 nmol, respectively with a RSD of 4.0 - 4.5%.

Paper Details

Date Published: 1 September 1991
PDF: 7 pages
Proc. SPIE 1510, Chemical and Medical Sensors, (1 September 1991); doi: 10.1117/12.47122
Show Author Affiliations
Loic J. Blum, Univ. Claude Bernard-Lyon 1 (France)
Sabine Gautier, Univ. Claude Bernard-Lyon 1 (France)
Pierre R. Coulet, Univ. Claude Bernard-Lyon 1 (France)

Published in SPIE Proceedings Vol. 1510:
Chemical and Medical Sensors
Otto S. Wolfbeis, Editor(s)

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