One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20)

Ting Wang, Takanori Aoki, Kazushi Iwata, Takashi Takata, Takashi Uchida, Vera Knäuper, Elena Llano, Yasunori Okada, John D. Bartlett

研究成果: Article査読

6 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)530-537
ページ数8
ジャーナルEuropean Journal of Oral Sciences
108
6
DOI
出版ステータスPublished - 2000 12

ASJC Scopus subject areas

  • 歯科学(全般)

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