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Fast Homogeneous Assay for Plasma Procarboxypeptidase U

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Published/Copyright: June 1, 2005
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Volume 39 Issue 9

Abstract

Carboxypeptidase U (EC 3.4.17.20, CPU, TAFIa) is a novel determinant of the fibrinolytic rate. It circulates as an inactive zymogen, procarboxypeptidase U, which becomes active during the process of coagulation. We developed a high throughput method on microtiter plates for the determination of the procarboxypeptidase U concentration in human plasma samples. Following activation of procarboxypeptidase U by thrombin-thrombomodulin, the resulting enzyme activity cleaves p-OH-Hip-Arg and the generated p-OH-hippuric acid is converted by hippuricase to p-hydroxybenzoic acid and glycine. Finally, oxidative coupling of p-hydroxybenzoic acid with 4-aminoantipyrine by NaIO4 forms the quinoneimine dye. The absorbance of the latter dye is determined at 506 nm in a microtiter plate reader. A mean value of 620 U/l was found, with a CV of 3.0% within-run and 4.3% between-run. The assay showed a good correlation with the activities observed using a HPLC assay as reference method (n = 25, r = 0.979). The presented method enables the routine analysis of large sample pools in clinical setting.

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Published Online: 2005-06-01
Published in Print: 2001-09-07

Copyright © 2001 by Walter de Gruyter GmbH & Co. KG

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  2. Herbert Keller January 16, 1925 June 25, 2001
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  4. Diagnostic Testing in Epstein-Barr Virus Infection
  5. Establishing a Reference System in Clinical Enzymology
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  7. Fast Homogeneous Assay for Plasma Procarboxypeptidase U
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https://doi.org/10.1515/CCLM.2001.133

Articles in the same Issue

  1. Joint Meeting of the German Society of Clinical Chemistry and of the German Society of Laboratory Medicine, Rostock, 1619 September 2001
  2. Herbert Keller January 16, 1925 June 25, 2001
  3. Routine Differential Diagnosis of Proteinurias by Capillary Electrophoresis
  4. Diagnostic Testing in Epstein-Barr Virus Infection
  5. Establishing a Reference System in Clinical Enzymology
  6. Prediction of the Development of Pregnancy-Induced Hypertensive Disorders in High-Risk Pregnant Women by Artificial Neural Networks
  7. Fast Homogeneous Assay for Plasma Procarboxypeptidase U
  8. Detection and Quantification of Small Numbers of Circulating Tumour Cells in Peripheral Blood Using Laser Scanning Cytometer (LSC®)
  9. Monotherapy with Metformin: Does It Improve Hypoxia in Type 2 Diabetic Patients?
  10. Cigarette Smoking and Hypertension Influence Nitric Oxide Release and Plasma Levels of Adhesion Molecules
  11. The Effect of Corticosteroids on Serum and Cerebrospinal Fluid Nitric Oxide Levels in Multiple Sclerosis
  12. Glutathione Transferase P1 Polymorphism in Neuroblastoma Studied by Endonuclease Restriction Mapping
  13. Biological Variation of Interleukin-1β, Interleukin-8 and Tumor Necrosis Factor-α in Serum of Healthy Individuals
  14. Reference Intervals and Variation for Urinary Epinephrine, Norepinephrine and Cortisol in Healthy Men and Women in Denmark
  15. Determination of Serum Cystatin C: Biological Variation and Reference Values
  16. Certification of the Mass Concentration of Creatine Kinase Isoenzyme 2 (CK-MB) in the Reference Material BCR 608
  17. Carbohydrate-Deficient Transferrin (CDT) Determination by Nephelometry Using a Commercial Kit. Analytical and Diagnostic Aspects
  18. Higher CK-MB Mass Values in Heparin Plasma than in Serum Measured with the Abbott AxSYM™ System
  19. Clinical Case Material for Teaching Clinical Chemistry and Laboratory Medicine
  20. IFCC News
  21. Meetings
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