ELISA for complexes between urokinase-type plasminogen activator and its receptor in lung cancer tissue extracts

Autor: Fred C.G.J. Sweep, Nils Brünner, H. de Witte, Niels Behrendt, Gunilla Høyer-Hansen, Theo Benraad, Keld Danø, Jan Grøndahl-Hansen, B Guldhammer-Skov, Helle Pappot
Rok vydání: 1997
Předmět:
Cancer Research
Pathology
medicine.medical_specialty
Lung Neoplasms
medicine.drug_class
Enzyme-Linked Immunosorbent Assay
Receptors
Cell Surface
Monoclonal antibody
Sensitivity and Specificity
Receptors
Urokinase Plasminogen Activator
Mice
Development of assays for prognostic factors in oncological endocrinology
medicine
Animals
Humans
skin and connective tissue diseases
neoplasms
GeneralLiterature_REFERENCE(e.g.
dictionaries
encyclopedias
glossaries)
Immunosorbent Techniques
Urokinase
medicine.diagnostic_test
biology
Antibodies
Monoclonal
Molecular biology
Urokinase-Type Plasminogen Activator
biological factors
Urokinase receptor
enzymes and coenzymes (carbohydrates)
Cross-Linking Reagents
Oncology
Epidermoid carcinoma
Polyclonal antibodies
Immunoassay
Ontwikkeling van meetmethodes voor prognostische factoren in de oncologische endocrinologie
biology.protein
Carcinoma
Squamous Cell
Rabbits
biological phenomena
cell phenomena
and immunity
Antibody
Plasminogen activator
medicine.drug
Zdroj: International Journal of Cancer, 72, pp. 416-423
International Journal of Cancer, 72, 416-423
International Journal of Cancer, 72, 3, pp. 416-423
ISSN: 0020-7136
Popis: A sandwich-type ELISA has been developed for the assessment of complexes between urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in extracts of squamous cell lung carcinomas. The assay is based on a combination of rabbit polyclonal anti-uPA antibodies and a biotinylated mouse anti-uPAR monoclonal antibody (MAb). The detection limit of the assay is approximately 0.5 fmol/ml. A linear dose-response is obtained with up to 40 fmol/ml of uPA:uPAR complexes, while uPA and uPAR separately do not cause any response in the ELISA. A buffer which has been used previously for optimal extraction of uPAR yields the highest amounts of uPA:uPAR complexes. Absorption of tumor extracts with anti-uPA or anti-uPAR MAbs results in a complete disappearance of the ELISA signal, demonstrating the specificity of the ELISA. The recovery of chemically cross-linked uPA:uPAR complexes added to tumor extracts varies between 80% and 105%. The intra- and inter-assay variation coefficients are 5.3% and 9.8%, respectively. Furthermore, a peptide antagonist for uPAR was employed to evaluate de novo uPA:uPAR complex formation during tumor tissue extraction and the immunoassay procedure. Our results strongly indicate that de novo complex formation is a major factor to consider and that complexes analyzed in the presence of this antagonist represent original uPA:uPAR complexes present prior to tumor tissue processing. The present ELISA appears suitable for studying the potential prognostic impact of uPA:uPAR complexes in lung tumor tissue as well as other types of cancer.
Databáze: OpenAIRE
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