Quantitation of the receptor for urokinase plasminogen activator by enzyme-linked immunosorbent assay.
Binding of the urokinase plasminogen activator (uPA) to a specific cell surface receptor (uPAR) plays a crucial role in proteolysis during tissue remodelling and cancer invasion. An immunosorbent assay for the quantitation of uPAR has now been developed. This assay is based on two monoclonal antibodies recognizing the non-ligand binding part of this receptor, and it detects both free and occupied uPAR, in contrast to ligand-binding assays used previously. In a variant of the assay, the occupied fraction of uPAR is selectively detected with a uPA antibody. To be used as a standard, a soluble variant of uPAR, suPAR, has been constructed by recombinant technique and the protein content of a purified suPAR standard preparation was determined by amino acid composition analysis. The sensitivity of the assay (0.6 ng uPAR/ml) is strong enough to measure uPAR in extracts of cultured cells and cancer tissue. Recent studies have shown that a high uPA level in tumor extracts is in some cancers associated with poor prognosis. The present assay will now allow similar prognostic studies of uPAR levels.
Rønne
E
E
Finsen Laboratory, Rigshospitalet, Copenhagen, Denmark.
Behrendt
N
N
Ploug
M
M
Nielsen
H J
HJ
Wöllisch
E
E
Weidle
U
U
Danø
K
K
Høyer-Hansen
G
G
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D016466:CHO Cells; D002277:Carcinoma / Q000737:chemistry; D003110:Colonic Neoplasms / Q000737:chemistry; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D051379:Mice; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D000911:Antibodies, Monoclonal; C524898:PLAUR protein, human; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins