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

I. Azumano, N. Fujimoto, M. Teshima, S. Yoshida, K. Iwata, Y. Okada

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Abstract

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.

Original languageEnglish
Pages (from-to)23-28
Number of pages6
JournalConnective Tissue
Volume30
Issue number1
Publication statusPublished - 1998 Jan 1

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Keywords

  • Matrilysin
  • Matrix metalloproteinase 7
  • Monoclonal antibody
  • Sandwich enzyme immunoassay
  • Tissue inhibitor of metalloproteinases

ASJC Scopus subject areas

  • Rheumatology

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