Design of coupled reactions for simplification of bioluminescence analysis.

Luminescence analysis may be applied to many substances by arranging a prior reaction producing a species entering the light-emitting reaction. Under favourable conditions the two consecutive reactions are carried out simultaneously as a one-step procedure. In a bioluminescence assay, luciferase stability is frequently a problem, making it desirable to develop analytical schemes where the analytical response becomes largely independent of any impaired luciferase activity. The value of maximal emission or an approached steady-state level is a convenient and usually well-defined analytical parameter. When recording this level it is important to design the participating reactions in a way that compensates for changes in luciferase activity.