Premium
One‐step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP‐20)
Author(s) -
Wang Ting,
Aoki Takanori,
Iwata Kazushi,
Takata Takashi,
Uchida Takashi,
Knäuper Vera,
Llano Elena,
Okada Yasunori,
Bartlett John D.
Publication year - 2000
Publication title -
european journal of oral sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.802
H-Index - 93
eISSN - 1600-0722
pISSN - 0909-8836
DOI - 10.1034/j.1600-0722.2000.00922.x
Subject(s) - monoclonal antibody , immunoassay , chinese hamster ovary cell , microbiology and biotechnology , recombinant dna , chemistry , saliva , matrix metalloproteinase , matrix (chemical analysis) , peroxidase , enzyme , antibody , chromatography , biology , biochemistry , immunology , receptor , gene
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.