Document Detail


Newer synthetic peptide substrates in coagulation testing: some practical considerations for automated methods.
MedLine Citation:
PMID:  6658459     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
More than 100 chromogenic and fluorogenic peptide substrates are now available for the evaluation of coagulation and related parameters. Many of these substrates exhibit undesirable physical properties, such as insolubility, surface adsorption, and interaction with endogenous plasma proteins. Some of these substrates are capable of inhibiting serine protease generation during activation in the global assay. In order to develop synthetic chromogenic substrates with desirable physical and biochemical characteristics, modified amino acids, such as CHG, CHT, and Nleu, have been utilized. Similarly, to provide a favorable molecular environment to facilitate enzyme and synthetic substrate interactions, various molecular manipulations, such as the introduction of bulky groups, is helpful in developing substrates for protein Ca and C1-esterase. Substrates for Factor Xa, CH3-O-CO-CHG-Arg-pNA (bovine Xa, Km 2.5 X 10(-4) M; human, Km 3.5 X 10(-4) M); thrombin, H-D-CHT-Ala-Arg-pNA (bovine thrombin, Km 3 X 10(-6) M; human thrombin, Km 6 X 10(-6) M); plasmin, H-D-Nleu-CHT-Lys-pNA (human plasmin, Km 2.2 X 10(-5) M) were found to have identical or superior biochemical characteristics to the earlier substrates. These newer substrates were found to be more soluble (greater than 5 X 10(-4) M) in physiologic buffer, less susceptible to autoamidolysis at reaction conditions, and did not produce opacity of the test solution in final concentrations of 5 X 10(-4) M. Comparable results on normal and pathologic plasma samples were obtained in various laboratory assays that utilize currently available substrates for Factors Xa and IIa, kallikrein, and plasmin (R = greater than 0.9). We propose that prior to the application of a new synthetic substrate in a given assay, a careful biochemical and physical screening of the substrate, the assay conditions, and the interaction of substrates with plasma proteins is highly desirable.
Authors:
L G Svendsen; J Fareed; J M Walenga; D Hoppensteadt
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Seminars in thrombosis and hemostasis     Volume:  9     ISSN:  0094-6176     ISO Abbreviation:  Semin. Thromb. Hemost.     Publication Date:  1983 Oct 
Date Detail:
Created Date:  1984-02-14     Completed Date:  1984-02-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0431155     Medline TA:  Semin Thromb Hemost     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  250-62     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Blood Coagulation Factors / analysis*
Blood Coagulation Tests / instrumentation,  methods*
Chromogenic Compounds
Female
Fluorescent Dyes
Humans
Male
Peptides
Substrate Specificity
Chemical
Reg. No./Substance:
0/Blood Coagulation Factors; 0/Chromogenic Compounds; 0/Fluorescent Dyes; 0/Peptides

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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