Bioluminescence assays with immobilised bacterial luciferase using flow injection analysis

Abstract

A flow injection manifold incorporating bacterial luciferase and oxidoreductase (from Vibrio harveyi) co-immobilised on cyanogen bromide-activated Sepharose 4B is described. The protein recovery was 86 and 96% for luciferase and oxidoreductase, respectively, and the activity recovery was 14% for luciferase and 33% for oxidoreductase. The activity was retained for several days at room temperature. Calibration data for NADH, NADPH, FMN and FMNH₂ over the range 1.0 × 10^–9–1.0 × 10^–4M is given for the optimised system.