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Measurement of urokinase-type plasminogen activator (u-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies.
Thromb Haemost. 1986 Dec 15; 56(3):411-4.TH

Abstract

An enzyme-linked immunosorbent assay (ELISA) for the measurement of urokinase-type plasminogen activator (u-PA) was developed. Three murine monoclonal antibodies to single chain urokinase-type plasminogen activator (scu-PA) were isolated and shown to react with non-overlapping epitopes in scu-PA. Two of the three antibodies were coated onto microtiter plates and bound u-PA was quantitated with the third antibody conjugated to horseradish peroxidase. The assay was equally sensitive to Mr 54,000 u-PA in the single chain or two-chain form but did not respond to Mr 33,000 urokinase. The lower limit of sensitivity of the assay was 0.1 ng/ml in buffer and 1 ng/ml in plasma. Coefficients of variation of the assay at physiological levels of u-PA were 6.5 percent within assays and 13 percent between assays. The level of u-PA in normal resting plasma was 1.9 +/- 0.66 ng/ml (mean +/- SD, n = 54). The assay can be performed within one working day and provides an efficient, reproducible, and stable means for the measurement of u-PA in biological fluids. As such it may facilitate physiological and pharmacological studies of urokinase-type plasminogen activators in man.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

3105111

Citation

Darras, V, et al. "Measurement of Urokinase-type Plasminogen Activator (u-PA) With an Enzyme-linked Immunosorbent Assay (ELISA) Based On Three Murine Monoclonal Antibodies." Thrombosis and Haemostasis, vol. 56, no. 3, 1986, pp. 411-4.
Darras V, Thienpont M, Stump DC, et al. Measurement of urokinase-type plasminogen activator (u-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies. Thromb Haemost. 1986;56(3):411-4.
Darras, V., Thienpont, M., Stump, D. C., & Collen, D. (1986). Measurement of urokinase-type plasminogen activator (u-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies. Thrombosis and Haemostasis, 56(3), 411-4.
Darras V, et al. Measurement of Urokinase-type Plasminogen Activator (u-PA) With an Enzyme-linked Immunosorbent Assay (ELISA) Based On Three Murine Monoclonal Antibodies. Thromb Haemost. 1986 Dec 15;56(3):411-4. PubMed PMID: 3105111.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Measurement of urokinase-type plasminogen activator (u-PA) with an enzyme-linked immunosorbent assay (ELISA) based on three murine monoclonal antibodies. AU - Darras,V, AU - Thienpont,M, AU - Stump,D C, AU - Collen,D, PY - 1986/12/15/pubmed PY - 1986/12/15/medline PY - 1986/12/15/entrez SP - 411 EP - 4 JF - Thrombosis and haemostasis JO - Thromb Haemost VL - 56 IS - 3 N2 - An enzyme-linked immunosorbent assay (ELISA) for the measurement of urokinase-type plasminogen activator (u-PA) was developed. Three murine monoclonal antibodies to single chain urokinase-type plasminogen activator (scu-PA) were isolated and shown to react with non-overlapping epitopes in scu-PA. Two of the three antibodies were coated onto microtiter plates and bound u-PA was quantitated with the third antibody conjugated to horseradish peroxidase. The assay was equally sensitive to Mr 54,000 u-PA in the single chain or two-chain form but did not respond to Mr 33,000 urokinase. The lower limit of sensitivity of the assay was 0.1 ng/ml in buffer and 1 ng/ml in plasma. Coefficients of variation of the assay at physiological levels of u-PA were 6.5 percent within assays and 13 percent between assays. The level of u-PA in normal resting plasma was 1.9 +/- 0.66 ng/ml (mean +/- SD, n = 54). The assay can be performed within one working day and provides an efficient, reproducible, and stable means for the measurement of u-PA in biological fluids. As such it may facilitate physiological and pharmacological studies of urokinase-type plasminogen activators in man. SN - 0340-6245 UR - https://www.unboundmedicine.com/medline/citation/3105111/Measurement_of_urokinase_type_plasminogen_activator__u_PA__with_an_enzyme_linked_immunosorbent_assay__ELISA__based_on_three_murine_monoclonal_antibodies_ L2 - https://antibodies.cancer.gov/detail/CPTC-HLA-B-1 DB - PRIME DP - Unbound Medicine ER -