Abstract
The activity of matrix metalloproteinases (MMPs) is closely regulated by tissue inhibitors of metalloproteinases (TIMPs) under pathophysiological conditions. The quantitative imbalance between MMPs and TIMPs in tissues and body fluids is considered to cause tumor invasion, metastasis, and various connective tissue diseases such as arthritis and periodontitis (1). MMPs and TIMPs have been identified and purified from various samples such as cell-conditioned media, tissue homogenates, sera, and so on. Furthermore, monoclonal antibodies against MMPs and TIMPs have been also prepared, and enzyme immunoassays (EIAs), which can quantitate MMPs and TIMPs in specimens, have been established. A two-step sandwich EIA for human MMP-3 was prepared using the combination of a monoclonal antibody as the solid-phase with rabbit polyclonal antibody and horseradish peroxidase (HRP)-labeled donkey antirabbit gamma globulin as the second phase (2). EIAs for human MMP-9 were developed using MMP-9 specific polyclonal antibody (3,4) or monoclonal antibodies (5). For quantitative determination of MMP-8, a two-step sandwich EIA was developed using polyclonal antibody (6). We also established one-step sandwich EIAs for MMPs and TIMPs using two kinds of monoclonal antibodies (7–16).
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References
Birkedal-Hansen H., Moore W. G. I., Bodden M. K., Windsor L. J., Birkedal-Hansen B., Decarlo A., and Engler J. A. (1993) Matrix metalloproteinases: a review. Oral Biol. Med.. 4, 197–250.
Cooksley S., Hipkiss J. B., Tickle S. P., Holmes-Ievers E., Docherty A. J. P., Murphy G., and Lawson A. D. G. (1990) Immunoassays for the detection of human collagenase, stromelysin, tissue inhibitor of metalloproteinases (TIMP) and enzyme-inhibitor complex. Matrix. 10, 285–291.
Ballin M., Gomez D. E., Sinha C. C., and Thorgeirsson U. P. (1988) Ras oncogene mediated induction of a 92 kDa metalloproteinase: strong correlation with the malignant phenotype. Biochem. Biophys. Res. Commun.. 154, 832–838.
Kjeldsen L., Bjerrum O. W., Hovgaard D., Johnson A. H., Sehested M., and Borregaard H. (1992) Human neutrophil gelatinase: a marker for circulating blood neutrophils. Purification and quantitation by enzyme linked immunosorbent assay. Eur. J. Haematol.. 49, 180–191.
Zucker S., Lysik R. M., Zarrabi M. H., and Moll U. (1993) Mr 92,000 type IV collagenase is increased in plasma of patients with colon cancer and breast cancer. Cancer Res.. 53, 140–146.
Bergmann U., Michaelis J., Oberhoff R., Knäuper V., Beckmann R., and Tscheshe H. (1989) Enzyme linked immunosorbent assay (ELISA) for the quantitative determination of human leukocyte collagenase and gelatinase. J. Clin. Chem. Clin. Biochem.. 27, 351–359.
Zhang J., Fujimoto N., Iwata K., Sakai T., Okada Y., and Hayakawa T. (1993) A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 1 (interstitial collagenase) using monoclonal antibodies. Clin. Chim. Acta.. 219, 1–14.
Fujimoto N., Mouri N., Iwata K., Ohuchi E., Okada Y., and Hayakawa T. (1993) A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 2 (72-kDa gelatinase/type IV collagenase) using monoclonal antibodies. Clin. Chim. Acta.. 221, 91–103.
Obata K., Iwata K., Okada Y., Kohrin Y., Ohuchi E., Yoshida S., Shinmei M., and Hayakawa T. (1992) A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 3 (stromelysin-1) using monoclonal antibodies. Clin. Chim. Acta.. 211, 59–72.
Ohuchi E., Azumano I., Yoshida S., Iwata K., and Okada Y. (1996) A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies. Clin. Chim. Acta.. 244, 181–198.
Matsuki H., Fujimoto N., Iwata K., Knäuper V. Okada Y., and Hayakawa T.(1996) A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 8 (neutrophil collagenase) using monoclonal antibodies. Clin. Chim. Acta.. 244, 129–143.
Fujimoto N., Hosokawa N., Iwata K., Shinya T., Okada Y., and Hayakawa T. (1994) A one-step sandwich enzyme immunoassay for inactive precursor and complexed forms of human matrix metalloproteinase 9 (92 kDa gelatinase/type IV collagenase, gelatinase B) using monoclonal antibodies. Clin. Chim. Acta.. 231, 79–88.
Tamei H., Azumano I., Iwata K., Yoshihara Y., López-Otín C., Vizoso F., Knäuper V., and Murphy G. (1998) One-step sandwich enzyme immunoassays for human matrix metalloproteinase 13 (collagenase-3) using monoclonal antibodies. Connect. Tissue. 30, 15–22.
Kodama S., Iwata K., Iwata H., Yamashita K., and Hayakawa T. (1990) Rapid one-step sandwich enzyme immunoassay for tissue inhibitor of metallo-proteinases: an application of rheumatoid arthritis serum and plasma. J. Immunol. Methods. 127, 103–108.
Fujimoto N., Zhang J., Iwata K., Shinya T., Okada Y., and Hayakawa T.(1993) A one-step sandwich enzyme immunoassay for tissue inhibitor of metalloproteinase-2 using monoclonal antibodies. Clin. Chim. Acta.. 220, 31–45.
Kodama S., Kishi J., Obata K., Iwata K., and Hayakawa T. (1987) Monoclonal antibodies to bovine collagenase inhibitor. Col. Rel. Res.. 7, 341–350.
Ishikawa E., Imagawa M., Hashida S., Yoshitake S., Hamaguchi Y., and Ueno T. (1983) Enzyme-labeling of antibodies and their fragments for enzyme immunoassay and immunohistochemical staining. J. Immunoassay. 4, 209–327.
Hashida S., Imagawa M., Inoue S., Ruan K.-h., and Ishikawa E. (1984) More useful maleimide compounds for the conjugation of Fab′ to horseradish peroxidase through thiol group in the hinge. J. Appl. Biochem.. 6, 56–63.
LaemmLi U. K. and Favre M. (1973) Maturation of the head of bacteriophage T4. 1. DNA packaging events. J. Mol. Biol.. 80, 597–599.
Cooper T. W., Eisen A. Z., Stricklin G. P., and Welgus H. G. (1985) Platelet derived collagenase inhibitor: characterization and subcellular localization. Proc. atl. Acad. Sci. USA. 82, 2779–2783.
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© 2001 Humana Press Inc.
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Fujimoto, N., Iwata, K. (2001). Use of EIA to Measure MMPs and TIMPs. In: Clark, I.M. (eds) Matrix Metalloproteinase Protocols. Methods in Molecular Biology™, vol 151. Humana Press. https://doi.org/10.1385/1-59259-046-2:347
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DOI: https://doi.org/10.1385/1-59259-046-2:347
Publisher Name: Humana Press
Print ISBN: 978-0-89603-733-5
Online ISBN: 978-1-59259-046-9
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