In vitro inhibition of human MMP-2 collagenolytic and gelatinolytic activity by neutralizing monoclonal antibodies.
Matrix metalloproteinases (MMPs) have been implicated in the degradation of the extracellular matrix in normal and pathological tissue remodelling. Among the MMPs, MMP-2 is the most commonly studied protease that has been involved in cancer, inflammation, infective diseases, degenerative diseases of the brain and vascular diseases. In this study, monoclonal antibodies (MAbs) were generated against human MMP-2, purified, characterized and tested for their ability to inhibit the enzymatic activity of MMP-2. Out of 12 positive clones generated against MMP-2, 2 clones (F2-1-11 and G8-25-5) were selected for further characterization. The selected clones react specifically with human pro and active form of MMP-2 in enzyme linked immunosorbant assay (ELISA), dot immunobinding assay (DIA) and Western blot and do not cross react with other human metalloproteinases or MMP-2 from other species. Additionally, these MAbs (F2-1-11 and G8-25-5) selectively inhibit collagenolytic and gelatinolytic activity of APMA ((p-aminophenylmercuric acetate)-activated-pro-MMP-2 and MMP-2, respectively.
Emara
Marwan
M
Department of Oncology, University of Alberta, Edmonton, AB, Canada. memara@ualberta.ca
Wozniak
Mietek
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D017364:Collagenases / Q000276:immunology / Q000378:metabolism; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D018093:Gelatinases / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000276:immunology / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D061965:Matrix Metalloproteinase Inhibitors; D017364:Collagenases; D018093:Gelatinases; D020778:Matrix Metalloproteinase 2