Projects / Programmes
The production of human recombinant metalloproteinase inhibitor TIMP-1 and development of specific immunodiagnostic reagents for determination of the native TIMP-1 in biological samples.
| Code |
Science |
Field |
Subfield |
| 1.09.00 |
Natural sciences and mathematics |
Pharmacy |
|
| Code |
Science |
Field |
| B740 |
Biomedical sciences |
Pharmacological sciences, pharmacognosy, pharmacy, toxicology |
| T490 |
Technological sciences |
Biotechnology |
recombinant proteins, tissue inhibitor of metalloproteinases-1 (TIMP-1); Pichia pastoris, Echerichia coli; phage display; single-chain Fv fragments; fusion proteins; alkaline phosphatase; ELISA, melanoma, rheumatoid arthritis
Researchers (8)
Organisations (2)
Abstract
In normal physiological conditions high concentrations of unbound metalloproteinase inhibitors can only be found in immuno-priviledged tissues, constitutively lacking blood vessels, like cartilage and cornea. For angiogenesis to take place unhindered action of metalloproteinases is needed. Altered patterns of certain components of tissue metalloproteinases and their inhibitors have been observed in patients suffering from multiple sclerosis, cronical liver diseases, scleroderma and different types of cancers, like metastatic melanoma, glioma, lipoma, ovarian adenocarcinoma and colon carcinoma. In most cases a switch to malign phenotype in cancers is being accompanied by increased production of metalloproteinases in affected tissues, leading to neoangiogenesis and metastasing of tumors. To compensate for this imbalance the host organism starts to overproduce their inhibitors, one of them being also TIMP-1.Our aim is to prepare a higly purified human recombinant TIMP-1 by using bacterial (E. coli) as well as fungal (Pichia pastoris) expression systems. We will test TIMP-1 activity in different in vitro models to evaluate its potency as well as to explore the possibility for its potential therapeutic use. Once obtained, the recombinant TIMP-1 will also be used as a template for preparing completely recombinant immunodiagnostical reagents, to be integrated in a specific ELISA test constructed for determination of TIMP-1 levels in biological samples.In this way we are planning to measure and compare the amounts of TIMP-1 in serum samples of healthy volounteers, melanoma and rheumatoid arthrytis patients. With results obtained we are expecting to prove the prognostic value of increased TIMP-1 serum levels in tracking the disease development and also to unequivocally point out a great potential of recombinant antibody fragment technology in constructing different in vitro diagnostical tests.
