The use of substrates, the fluorescence properties from those of the converted products differ from each other, a method for the determination of the enzyme activity.

摘要

Method in demonstrating the presence of enzyme activity with the aid of a fluorogenic substrate (A) which is converted by the enzyme to a product (B) differing from (A) in respect of its fluorescent properties. The characteristic feature resides in that (A) and/or (B) carries a chromophore which is a triplet sensitizer having a triplet energy level above the excitation level of a lanthanide ion selected from among Eu?3+, Tb3+, Dy3+ and Sm3+¿ and which is capable of chelating said lanthanide ion by means of an oxygen and/or nitrogen atom in said chromophore, optionally together with chelating groups bound to said chromophore; the said enzymatic conversion from (A) to (B) resulting in that the said chelating capacity is either caused to disappear or created and/or the chromophore structure is caused to undergo a change; and measurement of fluorescence is performed in the presence of said lanthanide ion by means of time resolved fluorescence spectrometry.