| Collagenases contains signal sequence, propeptide, catalytic domain, fibronectin-like domain and hemopexin domain. It associates with healing, reproduction and involution of reproductive organs. It was found that collagenases were related with migration and metastasis of tumor, autoimmune diseases, dental diseases and tumor angiogenesis. In our research, we established a fast, accurate, low-cost and high-throughput chromogenic assay for blood-plasma active collagenaseâ…£in order to evaluation the content of collagenaseâ…£in healthy blood-plasma.The assay was based on measurement of primary amines exposed as a result of succinylated gelatin hydrolysis. The optical density of each reaction was determined at 405 nM using a Sunrise microplate reader. Chromatographic and detection conditions were optimized and performance of the method was evaluated by recovery experiments and precision experiments. It was compared with ELISA. The levels of collagenaseâ…£in 112 health persons'plasma were determined and analysed by SPSS statistical software. We collected 86 leucocythemia patients'plasma while early onset, after first chemotherapy and second chemotherapy, then detected collagenaseâ…£of these plasma.The whole determing time was within 1.5h, the linear range of this method was 1.5~10 mg/L,and the minimum detection limit is 0.965 mg/L. It was correlated coefficiently with ELISA ( R2=0.999 7 , P<0.01 ) .The within-run CV<3.16%,and between-run CV <9.81%, and the recoveries for blood-plasma sample were in the range of 99.23%~107.59%. 95% double-side-restricted content of collagenaseâ…£in healthy plasma was 33.38~49.80 mg/L. The patients'collagenaseâ…£were higher than nomal,and becoming lower after chemotherapy.This chromogenic assay for blood-plasma active collagenaseâ…£was very successful in measurement of collagenaseâ…£in blood-plasma and established 95% double-side-restricted content of collagenaseâ…£in healthy plasma . |
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