Cancerforskningsfondet Af 1989

Fonden Til Fremme Af Eksperimentel Cancerforskning

Fonden Til Fremme Af Eksperimental Cancerforskning

Keld Dano

Lars S Nielsen

Vincent Ellis

Charles Pyke

Niels Behrendt

Nils Bruenner

Peter A Andreasen

Gunilla Hoyer-Hansen

Nils Brunner

Peter Andreasen

Ebbe Ronne

a monoclonal antibody which binds a human endothelial type plasminogen activator inhibitor (PAI-1) produced by dexamethasone-treated human HT-1080 fibrosarcoma cells may be used, inter alia, for determining PAI-1 protein abundance in tumor tissue or a sample of a body fluid. Measurements of this parameter may be useful in predicting the presence or metastasis of a tumor, or of predicting the progression of a known malignant tumor.

PAI-1 determination and use thereof

A monoclonal or polyclonal antibody directed against urokinase plasminogen activator receptor (u-PAR), or a subsequence, analogue or glycosylation variant thereof. Antibodies are disclosed which react with free u-PAR or with complexes between u-PA and u-PAR and which are capable of 1) catching u-PAR in ELISA, or 2) detecting u-PAR, e.g. in blotting, or 3) in radioimmunoprecipitation assay precipitate purified u-PAR in intact or fragment form, or 4) is useful for immunohistochemical detection of u-PAR, e.g. in immunostaining of cancer cells, such as in tissue sections at the invasive front, or 5) inhibits the binding of pro-u-PA and active u-PA and thereby inhibits cell surface plasminogen activation. Methods are disclosed 1) for detecting or quantifying u-PAR, 2) for targeting a diagnostic to a cell containing a u-PAR on the surface, 3) for preventing or counteracting proteolytic activity in a mammal. Methods for for selecting a substance suitable for inhibiting u-PA/u-PAR interaction, for preventing or counteracting localized proteolytical activity in a mammal, for inhibiting the invasion and/or metastasis comprise the use of the antibodies and of nude mice inoculated with human cancer cells which are known to invade and/or metastasize in mice and having a distinct color, f.x. obtained by means of an enzyme and a chromogenic substrate for the enzyme, the color being different from the cells of the mouse.

Urokinase-type plasminogen activator receptor antibodies

In a method of producing monoclonal antibodies against endothelial type plasminogen activator inhibitor and immunologically similar inhibitors myeloma cells are fused with antibody-producing cells obtained from mammals which have been immunized with said plasminogen activator inhibitor or with antibody-producing cells, which in vitro have been immunized with said plasminogen activator inhibitor and the hybridomas producing antibodies against the above mentioned inhibitor are selected. The antibody-producing cells are preferably spleen cells or lymph node cells, most preferably spleen cells, obtained from mice immunized with the above mentioned inhibitor. The monoclonal antibodies against endothelial type plasminogen activator inhibitor can be used for purification of plasminogen activator inhibitor, for removal of plasminogen activator inhibitor, from body fluids and other biological materials, for neutralization of the inhibitory activity of the plasminogen activator inhibitor and for detection, identification, immunostaining and quantification, e.g. by the ELISA technique, of plasminogen activator inhibitor in body fluids, normal or malignant cells and tissues, and other biological materials. Furthermore, as some of the monoclonal antibodies obtained bound complexes of urikinase type plasminogen activator with endothelial type inhibitor while others did not, the monoclonal antibodies obtained can be used for the quantification of free versus complex-bound inhibitor.

Plasminogen activator inhibitor monoclonal antibodies, hybridomas,

Growing community of inventors

Charlottenlund, Denmark

Keld Dano