A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies

Eiko Ohuchia, Isao Azumano, Shin'ichi Yoshida, Kazushi Iwata, Yasunori Okada

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 7 (MMP-7, matrilysin, EC 3.4.24.23) was established with a pair of monoclonal antibodies prepared against the zymogen of MMP-7 (proMMP-7) purified from CaR-1 human rectal carcinoma cells and against an oligopeptide corresponding to the C-terminal domain of human proMMP-7. ProMMP-7 in samples was allowed to simultaneously react with both solid-phase and peroxidase-labeled antibodies. Sensitivity of the EIA system in the presence of EDTA was 0.05 μg/l (1.5 pg/assay) and linearity was obtained between 0.156-10 μg/l (4.68-300 pg/assay). ProMMP-7 levels in human sera from healthy subjects were shown to be in the range of 10.7 ± 18.8 μg/l. Gel filtration analyses indicated that proMMP-7 makes polymers after treatment with EDTA and suggested that proMMP-7 exists as polymers in normal human sera.

Original languageEnglish
Pages (from-to)181-198
Number of pages18
JournalClinica Chimica Acta
Volume244
Issue number2
DOIs
Publication statusPublished - 1996 Jan 31

Fingerprint

Matrix Metalloproteinase 7
Matrix Metalloproteinases
Immunoenzyme Techniques
Enzyme Precursors
Monoclonal Antibodies
Enzymes
Edetic Acid
Assays
Polymers
Oligopeptides
Serum
Peroxidase
Gel Chromatography
Healthy Volunteers
Gels
Cells
Carcinoma
Antibodies

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies. / Ohuchia, Eiko; Azumano, Isao; Yoshida, Shin'ichi; Iwata, Kazushi; Okada, Yasunori.

In: Clinica Chimica Acta, Vol. 244, No. 2, 31.01.1996, p. 181-198.

Research output: Contribution to journalArticle

Ohuchia, Eiko ; Azumano, Isao ; Yoshida, Shin'ichi ; Iwata, Kazushi ; Okada, Yasunori. / A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies. In: Clinica Chimica Acta. 1996 ; Vol. 244, No. 2. pp. 181-198.
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