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Quantification of Tissue Inhibitor of Metalloproteinases 2 in Plasma from Healthy Donors and Cancer Patients
Author(s) -
Larsen M. B.,
Stephens R. W.,
Brünner N.,
Nielsen H. J.,
Engelholm L. H.,
Christensen I. J.,
StetlerStevenson W. G.,
HøyerHansen G.
Publication year - 2005
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.2005.01585.x
Subject(s) - ethylenediaminetetraacetic acid , polyclonal antibodies , chemistry , matrix metalloproteinase , antibody , monoclonal antibody , cancer , microbiology and biotechnology , medicine , immunology , chelation , biochemistry , biology , organic chemistry
Tissue inhibitor of metalloproteinases (TIMP)‐2 is a highly conserved molecule, which binds both active and latent matrix metalloproteinase (MMP)‐2. TIMP‐2 is also involved in the activation of MMP‐2 on the cell surface. A quantitative enzyme‐linked immunosorbent assay (ELISA) was established and optimized for measurement of TIMP‐2 in plasma. The capturing antibody in the ELISA was a monoclonal, while the detecting antibody was a chicken polyclonal antibody recognizing the native form of human TIMP‐2. The levels of TIMP‐2 were measured in ethylenediaminetetraacetic acid (EDTA) and citrate plasma from healthy donors. The median values were determined as 163 ng/ml ( n = 186) with a range of 109–253 ng/ml for EDTA plasma and 139 ng/ml ( n = 77) with a range of 95–223 ng/ml for citrate plasma. The TIMP‐2 concentration in citrate plasma from 15 patients with advanced, stage IV breast cancer had a median value of 160 ng/ml, only slightly higher but statistically distinguishable from the level found in citrate plasma from the healthy donors. In addition, the TIMP‐2 concentration in EDTA plasma from colorectal cancer patients revealed a significantly higher level in plasma from patients with Dukes stage A ( P = 0.01) compared with patients with more advanced Dukes stages.