Development of an enzyme-linked immunosorbent assay to measure total TIMP-1 (free TIMP-1 and TIMP-1 in combination with matrix-metalloproteinases) and measurement of TIMP 1 and CRP in serum

Clin Chim Acta. 1995 Sep 15;240(2):137-54. doi: 10.1016/0009-8981(95)06137-7.


A panel of six monoclonal antibodies (MAbs) was raised against purified human fibroblast tissue inhibitor of metalloproteinase-1 (TIMP-1) and characterised. All possible antibody pairs were tested for their suitability as capture and revealing antibodies in a two-site enzyme-linked immunosorbent assay (ELISA) to measure total TIMP-1 (both free TIMP-1 and TIMP-1 together with matrix metalloproteinases (MMPs)). Using the best combination of MAbs the assay was optimised. The sensitivity of detection of the assay was 1.4 ng/ml, and inter- and intra-assay coefficients of variation were between 10.4-13.7% and 8.8-9.7%, respectively. Dilution series of human cerebrospinal and synovial fluids, plasma and sera paralleled those of the TIMP-1 standard curve indicating that the immunoreactivity detected in these samples was authentic TIMP-1. TIMP-2 shows no detectable cross reactivity in this assay confirming that this ELISA is specific for TIMP-1. The levels of total TIMP-1 and collagenase were measured in conditioned medium from A2058 human melanoma cells cultured in the absence or presence of human recombinant interleukin-1 alpha (hrIL-1 alpha). Total TIMP-1 was also measured in serum samples with known C-reactive protein (CRP) (n = 100) and alpha 1 antichymotrypsin (ACT) (n = 52) concentrations; no correlation was found between TIMP-1 levels and either of these acute phase reactants although the levels of TIMP-1 were raised when compared to normal sera. This ELISA provides a rapid and convenient procedure for the quantitation of total TIMP-1 in human biological fluids and supernatants from cultured cell lines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / analysis
  • Antibodies, Monoclonal / biosynthesis
  • Biotin
  • C-Reactive Protein / analysis*
  • C-Reactive Protein / cerebrospinal fluid
  • Cells, Cultured
  • Cross Reactions
  • Culture Techniques
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Matrix Proteins / analysis
  • Fibroblasts / enzymology
  • Glycoproteins / analysis*
  • Glycoproteins / blood
  • Glycoproteins / cerebrospinal fluid
  • Humans
  • Immunoblotting
  • Metalloendopeptidases / analysis*
  • Metalloendopeptidases / blood
  • Metalloendopeptidases / cerebrospinal fluid
  • Synovial Fluid / enzymology
  • Tissue Inhibitor of Metalloproteinases
  • Tumor Cells, Cultured
  • alpha 1-Antichymotrypsin / analysis
  • alpha 1-Antichymotrypsin / blood


  • Antibodies, Monoclonal
  • Extracellular Matrix Proteins
  • Glycoproteins
  • Tissue Inhibitor of Metalloproteinases
  • alpha 1-Antichymotrypsin
  • Biotin
  • C-Reactive Protein
  • Metalloendopeptidases