Wang, Ting, Aoki, Takanori, Iwata, Kazushi, Takata, Takashi, Uchida, Takashi, Knauper, Vera  ORCID: https://orcid.org/0000-0002-3965-9924, Llano, Elena, Okada, Yasunori and Bartlett, John D.
2000.
One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20).
European Journal of Oral Sciences
108
(6)
, pp. 530-537.
10.1034/j.1600-0722.2000.00922.x
|
Abstract
A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5–160 ng/ml (125–8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.
| Item Type: | Article |
|---|---|
| Date Type: | Publication |
| Status: | Published |
| Schools: | Dentistry |
| Publisher: | Wiley |
| ISSN: | 1600-0722 |
| Last Modified: | 13 Sep 2023 10:44 |
| URI: | https://orca.cardiff.ac.uk/id/eprint/161650 |
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