A one-step sandwich enzyme immunoassay (EIA) system for human active matrix metalloproteinase 7 (MMP-7, matrilysin, EC 3. 4. 24. 23) was established with a monoclonal antibody conjugate that reacts specifically with the neoepitope, the N-terminal residues 78 - 82, of MMP-7 and another monoclonal antibody to the C-terminal domain (residues 239 - 253) of MMP-7 as the solid-phase. Active MMP-7 in samples was allowed to react simultaneously with both solid-phase and peroxidaselabeled antibodies. Sensitivity of the EIA system was 1.1 ng/ml (55 pg/assay), and linearity was obtained between 2.5 and 160 ng/ml (125 - 8000 pg/assay). The EIA system recognized the free form of active MMP-7 and its complexes with tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) with the same degree of immunoreactivity, but not the precursor or intermediate form of MMP-7. The newly developed one-step sandwich EIA system could measure active MMP-7 in culture medium conditioned by CaR-1 human rectal carcinoma cells and in normal human sera when standard active MMP-7 was added but active MMP-7 was undetectable in normal human serum by the assay system. These findings suggest that the concentration of total active MMP-7 is lower than 1.1 ng/ml in normal human serum.
|Number of pages||6|
|Publication status||Published - 1998 Jan 1|
- Matrix metalloproteinase 7
- Monoclonal antibody
- Sandwich enzyme immunoassay
- Tissue inhibitor of metalloproteinases
ASJC Scopus subject areas