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The measurement of free‐proteinase activity in inflammatory bowel disease mucosa

Recently evidence of proteolytic damage in IBD has been demostrated (Kjeldsen et al. 1995; Murch et al. 1993) but the proteases responsible for this tissue damage remain to be elucidated. Previously we have shown an increase in gelatinolytic activity in inflamed > noninvolved > control mucosal biopsies using substrate gel zymography which correlated with disease severity (Baugh et al. 1999). Neutrophil-derived MMP-9 was confirmed to be the major protease responsible for this increase but MMP-1, MMP-2 and a number of unidentified serine proteases were also detected. In order to determine whether proteases detected by zymography and Western blotting were active or inhibitor-bound free-enzyme analysis was carried out.