ALTERNATE DETERMINATION OF ATP AND NADH WITH A SINGLE BIOLUMINESCENCE-BASED FIBEROPTIC SENSOR

A novel approach in the design of biosensors is reported. It consists of the use of two different enzymatic systems co-immobilized on the same support allowing the alternate detection of two different analytes without changing the sensing element of the sensor. Based on this approach, a multifunctional bioluminescence fiber-optic sensor for the microdetermination of ATP and NADH has been developed. The firefly luciferase from Photinus pyralis has been successfully co-immobilized with the bacterial bioluminescent system from Vibrio fischeri on a preactivated polyamide membrane. When using such a bioactive matrix associated with a fiber-optic sensor, the alternate measurement of ATP or NADH could be performed with the same probe in the linear range 1 × 10-10 to 1 × 10-6 M and 1 × 10-9 to 1 × 10-6 M, respectively. The relative standard deviation was around 6% at 9 × 10-9 M ATP or at 4 × 10-8 M NADH. © 1990.