Flow injection fluorescence measurement of collagenase using a mini-bioreactor with immobilized collagen labelled with Lucifer Yellow

Abstract

A rapid, sensitive and reusable system has been developed for the analysis of collagenase. The system is based on flow injection of the enzyme into an immobilized Lucifer Yellow-labelled-collagen mini-bioreactor with fluorescence measurement. The injected collagenase digests the substrate within the bioreactor and releases fluoro-fragments, which are detected downstream by a fluorimeter. The relationship between the observed signal and the amount (0–100 ng) of collagenase injected is linear (r= 0.999). The response is fast (3 min response time) and reproducible (s_r= 3.6%).