دورية أكاديمية
Establishment of a standard assay method for human thrombomodulin and determination of the activity of the Japanese reference standard.
العنوان: | Establishment of a standard assay method for human thrombomodulin and determination of the activity of the Japanese reference standard. |
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المؤلفون: | Niimi S; Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan. niimi@nihs.go.jp, Oshizawa T, Naotsuka M, Ohba S, Yokozawa A, Murata T, Hayakawa T |
المصدر: | Biologicals : journal of the International Association of Biological Standardization [Biologicals] 2002 Mar; Vol. 30 (1), pp. 69-76. |
نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Academic Press Country of Publication: England NLM ID: 9004494 Publication Model: Print Cited Medium: Print ISSN: 1045-1056 (Print) Linking ISSN: 10451056 NLM ISO Abbreviation: Biologicals Subsets: MEDLINE |
أسماء مطبوعة: | Original Publication: London ; San Diego : Academic Press, c1990- |
مواضيع طبية MeSH: | Reference Standards*, Thrombomodulin/*analysis, Analysis of Variance ; Aniline Compounds/chemistry ; Chemistry, Clinical/standards ; Dose-Response Relationship, Drug ; Fibrinolytic Agents/chemistry ; Hot Temperature ; Humans ; Intercellular Signaling Peptides and Proteins ; Japan ; Laboratories/standards ; Peptides/pharmacology ; Temperature ; Thrombin/analysis ; Time Factors |
مستخلص: | This study was undertaken to establish a standard method for determination of the activity of human (h) thrombomodulin (TM). The reactions mainly consisted of formation of a h-TM and h-thrombin complex, activation of h-protein C by the complex and digestion of substrate by activated h-protein C. Linear time-dependent formation of p -nitroaniline from the substrate, S-2366, was observed up to 12 min during measurement of the activity of urinary h-TM (uh-TM) reference material by the standard method. Therefore, 10 minutes was established as the reaction time in the standard method. In the standard method, we defined the activity of h-TM forming 0.1 micromol of p -nitroaniline per min in the reaction as 1 JRS Unit. Recombinant h-TM (rh-TM) and uh-TM reference material gave rectilinear dose-response curves within a certain range of specific activities by their original methods in the standard method. The validity of the standard method was assessed based on the coefficients of variation (CV) obtained in the various measurements of h-TM. Intra-batch precision (CV) of h-thrombin and h-protein C was 2.90% and 6.57%, respectively, in the measurement of uh-TM activity. The intra-sample, inter-day, and inter-laboratory precision (CV) was 1.30%, 1.63% and 5.02%, respectively, in the measurement of the first Japanese reference standard for h-TM coded TJRS1. In these assays, the activity of the first Japanese standard was also determined and found to be 205 JRS units per ampoule. When the stability of the Japanese reference standard was assessed by measuring of the standard stored under various thermal conditions, the predicted loss of activity assuming monomolecular degradation according to the Arrhenius equation was less than 3.0% during 103 years at -20 degrees C. These results indicate that the standard method is appropriate for determination of the activity of h-TM and that the Japanese reference standard for h-TM, whose activity was determined here, can be stored at -20 degrees C for long periods without loss of activity. (Copyright 2002 The International Association for Biologicals.) |
المشرفين على المادة: | 0 (Aniline Compounds) 0 (Fibrinolytic Agents) 0 (Intercellular Signaling Peptides and Proteins) 0 (Peptides) 0 (Thrombomodulin) 0 (snake venom protein C activator) 1MRQ0QZG7G (4-nitroaniline) EC 3.4.21.5 (Thrombin) |
تواريخ الأحداث: | Date Created: 20020216 Date Completed: 20020513 Latest Revision: 20191210 |
رمز التحديث: | 20231215 |
DOI: | 10.1006/biol.2001.0318 |
PMID: | 11846431 |
قاعدة البيانات: | MEDLINE |
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