AN ENZYME LINKED IMMUNOSORPTION ASSAY FOR TISSUE FACTOR PATHWAY INHIBITOR
| Autor: | Helle Ørsted, Anita Svendsen, Per B. Øtergaard, John-Bjarne Hansen, Per Morten Sandset, Ole Nordfang, Thomas Christian Beck |
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| Rok vydání: | 1997 |
| Předmět: |
biology
T-plasminogen activator Lipoproteins Reproducibility of Results Enzyme-Linked Immunosorbent Assay Hematology Sensitivity and Specificity Molecular biology Tissue factor pathway inhibitor Fibrinolytic Agents Reference Values Enzyme inhibitor Polyclonal antibodies Blood plasma Monoclonal biology.protein Animals Humans Rabbits Reagent Kits Diagnostic Antibody Quantitative analysis (chemistry) Factor Xa Inhibitors |
| Zdroj: | Thrombosis Research. 87:447-459 |
| ISSN: | 0049-3848 |
| DOI: | 10.1016/s0049-3848(97)00161-8 |
| Popis: | An assay for the quantification of full-length and carboxy-terminus truncated tissue factor pathway inhibitor (TFPI) has been developed. The assay is a classical two-antibody sandwich assay with a monoclonal capture antibody directed against the third Kunitz-type domain of human TFPI and a polyclonal rabbit peroxidase-labelled anti-human TFPI detecting antibody. The assay is sensitive to full-length and carboxy-terminus truncated TFPI with intact third Kunitz-type domain, but not to two-domain TFPI. TFPI associated with lipoproteins is not or only sparsely detected and TFPI in complex with factor Xa only partially measured. The assay gives linear reference curves in the dose range of 5 to 100 ng/ml in a double logarithmic plot. The normal range assessed from analyses on citrated plasma from 81 normal human donors is 7.8 to 26.0 ng/ml (average ± 2 SD, log-normal distribution). There is no statistically significant difference between TFPI levels measured in 10 fasting and 10 non-fasting individuals. The reproducibility of the assay is about 5.6–5.9% (relative standard error) and the within-days and between-days reproducibilities are 4.7–5.1% and 5.9–8.5%, respectively. The assay is in very good agreement with a commercial ELISA assay recently marketed. A robust, reproducible and convenient ELISA assay for the determination of full-length and three-domain TFPI has been developed. © 1997 Elsevier Science Ltd |
| Databáze: | OpenAIRE |
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