Chromogenic Thrombin Generation Assay

The thrombin-specific chromogenic substrate Tosyl-Gly-Pro-Arg-p-nitroanilide (Chromozym-TH) was purchased from Sigma (USA).
For the thrombin generation test, the following reagents were used: thrombin-specific fluorogenic substrate Z-Gly-Gly-Arg-AMC (Bachem, Switzerland); 7-amino-4-methylcoumarin, СаСl₂, dimethyl sulfoxide (DMSO), and NaCl (Sigma-Aldrich, USA); thromboplastin (Renam, Russia); phosphatidylserine (from pig brain) and phosphatidylcholine (from egg yolk) produced by Avanti Polar Lipids Inc. (USA); and 4-(2-hydroxyethyl)-1-piperazine-2-ethanosulphonic acid (HEPES, Fisher Biotech, USA).
The calibrator for the determination of thrombin activity in the CAT test (ThrCal) was purchased from Thrombinoscope BV (the Netherlands) through Laborama (Russia).
Dabigatran (BIBR 953) was obtained from Selleckhem.com. HC-IOC-019s (C₆H₅-SO₂-O-C₆H₄(CH₃)-O-(CH₂)₂-NC₅H₄(NH₂)) was synthesized at the Institute of Organic Chemistry of Russian Academy of Sciences (Russia)33 (link). Pradaxa (dabigatran etexilate) (capsules 110 mg) was from Boehringer Ingelheim (Germany).

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Gribkova I.V., Lipets E.N., Rekhtina I.G., Bernakevich A.I., Ayusheev D.B., Ovsepyan R.A., Ataullakhanov F.I, & Sinauridze E.I. (2016). The modification of the thrombin generation test for the clinical assessment of dabigatran etexilate efficiency. Scientific Reports, 6, 29242.

Publication 2016

Z-Gly-Gly-Arg-AMC phosphatidylserine phosphatidylcholine

7 amino 4 methylcoumarin Acid Bibr 953 Brain Capsules Chromogenic substrate Chromozym th Dabigatran Dabigatran etexilate Dimethyl sulfoxide Egg yolk Fluorogenic substrate Gly pro arg Hepes Lipids Nacl Phosphatidylcholine Phosphatidylserine Piperazine Pradaxa Thrombin Thromboplastin Z gly gly arg amc

Corresponding Organization : National Research Center for Hematology Russian Academy of Medical Sciences

Other organizations : Center for Theoretical Problems of Physicochemical Pharmacology, Central Scientific Research Institute of Traumatology and Orthopedics, Dmitry Rogachev National Research Center of Pediatric Hematology, Oncology and Immunology, Ministry of Health of the Russian Federation, Lomonosov Moscow State University

Top 5 similar protocols

Variable analysis

independent variables

Dabigatran (BIBR 953)
HC-IOC-019s (C6H5-SO2-O-C6H4(CH3)-O-(CH2)2-NC5H4(NH2))

dependent variables

Thrombin activity

control variables

Tosyl-Gly-Pro-Arg-p-nitroanilide (Chromozym-TH)
Z-Gly-Gly-Arg-AMC (thrombin-specific fluorogenic substrate)
7-amino-4-methylcoumarin
CaCl2
Dimethyl sulfoxide (DMSO)
Thromboplastin
Phosphatidylserine (from pig brain)
Phosphatidylcholine (from egg yolk)
4-(2-hydroxyethyl)-1-piperazine-2-ethanosulphonic acid (HEPES)
ThrCal (calibrator for the determination of thrombin activity in the CAT test)
Pradaxa (dabigatran etexilate)

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