Systematic separation and purification of elastase, gelatinase (matrix metalloproteinase 9), and collagenase (matrix metalloproteinase 8) from polymorphonuclear leukocytes in dialyzers previously used by patients with renal failure

We developed a simple and effective method for the systematic separation an d purification of human polymorphonuclear leukocyte (PMN) proteinases, elas tase, gelatinase (matrix metalloproteinase 9, type IV collagenase), and col lagenase (matrix metalloproteinase 8), derived from the extracts of hollow fiber dialyzers that had been utilized in the treatment of patients with re nal failure. The fraction containing elastase was grossly separated from th at containing gelatinase and collagenase by heparin-Sepharose chromatograph y and purified in an aprotinin column. The remaining two enzymes were then separated using the gelatin-Sepharose column after gel chromatography follo wing ammonium sulfate precipitation. Gelatinase and collagenase were furthe r purified by gelatin-Sepharose chromatography as a latent form and by coll agen-Sepharose chromatography as an activated form. This novel method offer s procedural advantages over existing methods that separate PMNs from the w hole blood of volunteers for experimental research purposes. (C) 2001 Acade mic Press.