A one-step monoclonal antibody-based sandwich enzyme immunoassay for human active matrilysin(MMP-7)and its complexes with TIMP-1 and TIMP-2

概要

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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 peroxidase-labeled antibodies. Sensitivity of the EIA system was 1.1ng / ml(55pg / assay), and linearity was obtained between 2.5 and 160ng / ml(125-8000pg / 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 on-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 1.1ng / ml in normal human serum.
日本結合組織学会の論文
1998-03-25