A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies.
A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 7 (MMP-7, matrilysin, EC 3.4.24.23) was established with a pair of monoclonal antibodies prepared against the zymogen of MMP-7 (proMMP-7) purified from CaR-1 human rectal carcinoma cells and against an oligopeptide corresponding to the C-terminal domain of human proMMP-7. ProMMP-7 in samples was allowed to simultaneously react with both solid-phase and peroxidase-labeled antibodies. Sensitivity of the EIA system in the presence of EDTA was 0.05 microgram/l (1.5 pg/assay) and linearity was obtained between 0.156-10 micrograms/l (4.68-300 pg/assay). ProMMP-7 levels in human sera from healthy subjects were shown to be in the range of 10.7 +/- 18.8 micrograms/l. Gel filtration analyses indicated that proMMP-7 makes polymers after treatment with EDTA and suggested that proMMP-7 exists as polymers in normal human sera.
Ohuchi
E
E
Biopharmaceutical Department, Fuji Chemical Industries, Ltd., Toyama, Japan.
Azumano
I
I
Yoshida
S
S
Iwata
K
K
Okada
Y
Y
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002850:Chromatography, Gel; D003110:Colonic Neoplasms / Q000097:blood* / Q000201:enzymology / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000097:blood; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D020783:Matrix Metalloproteinase 7; D008666:Metalloendopeptidases / Q000097:blood*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009367:Neoplasm Staging; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D012004:Rectal Neoplasms; D012016:Reference Values; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D004792:Enzyme Precursors; D010446:Peptide Fragments; D008666:Metalloendopeptidases; C099583:promatrilysin; D020783:Matrix Metalloproteinase 7