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Clinical Chemistry 45: 1206-1213, 1999;
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(Clinical Chemistry. 1999;45:1206-1213.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Determination of the Complex between Urokinase and Its Type-1 Inhibitor in Plasma from Healthy Donors and Breast Cancer Patients

Anders N. Pedersen1,2,a, Nils Brünner1, Gunilla Høyer-Hansen1, Peter Hamer3, David Jarosz3, Birthe Larsen1, Hans Jørgen Nielsen4 and Ross W. Stephens1

1 The Finsen Laboratory, Rigshospitalet, Strandboulevarden 49, 2100 Copenhagen, Denmark.

2 Department of Oncology, Herlev Hospital, Herlev 2730, Denmark.

3 Oncogene Science Diagnostics, Cambridge, MA 02142-1168.

4 Department of Surgical Gastroenterology, Hvidovre Hospital, Hvidovre 2650, Denmark.
a Author for correspondence. Fax 45 35 38 54 50.

Background: The complex between urokinase (uPA) and its type-1 inhibitor (PAI-1) is formed exclusively from the active forms of these components; thus, the complex concentration in a biological sample may reflect the ongoing degree of plasminogen activation. Our aim was to establish an ELISA for specific quantification of the uPA:PAI-1 complex in plasma of healthy donors and breast cancer patients.

Methods: A kinetic sandwich format immunoassay was developed, validated, and applied to plasma from 19 advanced-stage breast cancer patients, 39 age-matched healthy women, and 31 men.

Results: The assay detection limit was <2 ng/L, and the detection of complex in plasma was validated using immunoabsorption, competition, and recovery tests. Eighteen cancer patients had a measurable complex concentration (median, 68 ng/L; range, <16 to 8700 ng/L), whereas for healthy females and males the median signal values were below the detection limit (median, <16 ng/L; range, <16 to 200 ng/L; P <0.0001). For patient plasma, a comparison with total uPA and PAI-1 showed that the complex represented a variable, minor fraction of the uPA and PAI-1 concentrations of each sample.

Conclusion: The reported ELISA enables detection of the uPA:PAI-1 complex in blood and, therefore, the evaluation of the complex as a prognostic marker in cancer.© 1999 American Association for Clinical Chemistry




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W. Qin, W. Zhu, C. Wagner-Mann, and E. R. Sauter
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A. N. Pedersen, I. J. Christensen, R. W. Stephens, P. Briand, H. T. Mouridsen, K. Danø, and N. Brünner;
The Complex between Urokinase and Its Type-1 Inhibitor in Primary Breast Cancer: Relation to Survival
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