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A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 8 (neutrophil collagenase) using monoclonal antibodies

Matsuki, Hirokazu, Fujimoto, Noboru, Iwata, Kazushi, Knauper, Vera ORCID: https://orcid.org/0000-0002-3965-9924, Okada, Yasunori and Hayakawa, Taro 1996. A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 8 (neutrophil collagenase) using monoclonal antibodies. Clinica Chimica Acta 244 (2) , pp. 129-143. 10.1016/0009-8981(95)06197-5

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Abstract

A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 8 (MMP-8, neutrophil collagenase, EC 3.4.24.7) has been established with a pair of monoclonal antibodies prepared against the zymogen of MMP-8 purified from human neutrophils. MMP-8 in samples simultaneously reacted with both solid-phase and peroxidaselabeled antibodies. Sensitivity of this EIA system was 0.34 μg/l (5.7 pg/assay) and linearity was obtained between 0.5 and 500 μg/l (8.3–8300 pg/assay). The EIA system recognized both precursor and active forms of MMP-8 but not MMP-8 complexed with tissue inhibitors of metalloproteinases. There was no difference in the MMP-8 levels between the plasma samples from patients with rheumatoid arthritis or osteoarthritis and those from healthy subjects (median 6.2 μg/l, range 1.5–28 μg/l). However, the level in synovial fluids from patients with rheumatoid arthritis (median 345 μg/l, range 84–2860 μg/l) was shown to be higher than that from osteoarthritic patients. MMP-8 levels in human whole saliva from patients with periodontal diseases (median 282 μg/l, range 0–1420 μg/l) were also significantly higher than those from clinically healthy subjects (median 25 μg/l, range 0–100 μg/l). Immunoreactivity analyses showed that MMP-8 species in normal human plasma exists as a precursor but not as a complex form with tissue inhibitor of metalloproteinases (TIMP)-1 or TIMP-2.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Dentistry
Publisher: Elsevier
ISSN: 0009-8981
Date of Acceptance: 11 October 1995
Last Modified: 14 Sep 2023 12:44
URI: https://orca.cardiff.ac.uk/id/eprint/161623

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