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Measurement of the noncomplexed free fraction of tissue inhibitor of metalloproteinases I in plasma by immunoassay

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Publisher
AMER ASSOC CLINICAL CHEMISTRY
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Keywords
  • Medicine And Health Sciences
Disciplines
  • Biology

Abstract

Background: We previously found differences in total concentrations of tissue inhibitor of metalloproteinases 1 (TIMP-1) in plasma from donors and cancer patients. Because TIMP-1 can exist in more than one molecular form, a new immunoassay to specifically detect free TIMP-1 was developed and concentrations were determined in plasma from healthy donors and colorectal cancer (CRC) patients. Methods: We established and validated an immunoassay for the specific measurement of free TIMP-1 that uses a polyclonal anti-TIMP-1 antibody for capture and a monoclonal anti-TIMP-1 antibody that binds only free TIMP-1 for detection of antigen. Plasma samples from healthy donors and CRC patients were assayed for free TIMP-1. Total TIMP-1 was measured by our previously published assay. Results: The mean (SD) concentrations of free TIMP-1 were similar in citrate 55.5 (11.5) mug/L and EDTA plasma 58.9 (13.3) mug/L from 76 donors (r(2) = 0.82). In 154 donors, the ratio of free TIMP-1 mean (SD), 64.5 (18.0) mug/L to total TIMP-1 83.8 (19.8) mug/L in EDTA plasma was 0.77. Plasma concentrations of free and total TIMP-1 correlated significantly to age (free, r(2) = 0.19; total, r(2) = .0.27; P < 0.0001), increasing 50% over an age span of 45 years. Free and total TIMP-1 were significantly increased in CRC patients (P < 0.0001), whereas the ratio of free to total TIMP-1 (mean, 0.58) was significantly lower than in donors. Conclusions: Most of the TIMP-1 in donor plasma is present in its free form, and free TIMP-1 increases with age. Free and total TIMP-1 are increased in CRC patient plasma, but the ratio of free to total TIMP-1 is significantly lower in these patients than in donors. (C) 2002 American Association for Clinical Chemistry.

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