Title
A novel and simple assay for determination of gelatinase-b (mmp-9) activities in biological fluids
Author
Gaubius Instituut TNO
Verheijen, J.
Visser, H.
Konttinen, Y.
Verspaget, H.
Duffy, J.
Hanemaaijer, R.
Publication year
1997
Abstract
Here we describe a new principle to assess specifically the activity of the different members of the human matrix-metalloproteinase family (MMPs) by a colorimetric assay. Using protein engineering a modified pro-urokinase has been made in which the activation sequence, normally recognized by plasmin (PRFK IIGG) has been replaced by a sequence that will be recognized by MMPs (RPLG IIGG) The active urokinase resulting from the activation of modified pro-urokinase by MMPs can be measured directly using a specific chromogenic peptide substrate for urokinase. Next to overall collagenase activity the assay can be made specific for the various MMPs using monoclonal antibodies and/or specific sites of hydrolysis. Following catching by specific antibodies of a particular MMP from biological fluids or tissue culture media, MMP-activity of both active and 'to be activated' MMP can be analyzed. Due to the catching no side-effects of disturbing factors in the biological fluids will interfere with the assay. Using this assay a significant increased level of both active and latent MMP-9 could be measured in saliva from patients with Sjögren syndrome compared with normals (2.46 vs 0.8 p
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DOI
https://doi.org/10.1016/s0268-9499(97)80262-7
TNO identifier
234226
ISSN
1369-0191
Source
Fibrinolysis and Proteolysis, 11 (11), 41
Document type
article