A drug-specific OSA calibration curve was constructed starting from 1.5 IU/mL (labeled concentration based on the manufacturer’s certificate of potency) of each EHL-rFVIII product, as calibrator, using HemosIL FVIII Deficient Plasma (PC8) and the SynthASil reagent (both from Werfen). Preliminary measurements showed that using the specific EHL-rFVIII product calibration curve, the use of SynthAsil or SynthaFax as activator did not provide different results for the measurement of EHL-rFVIII level in samples prepared with the same nominal FVIII concentration. Hence, we decided to use only the SynthASil reagent to carry out EHL-rFVIII-calibrated OSA. All EHL products’ levels were measured on serial dilutions for the 2 concentration ranges (ie, 1.0/0.5/0.1/0.05/0.025 and 0.1/0.05/0.025/0.0125/0.006125 IU/mL) by using the specific EHL-rFVIII-based calibration curve as a reference.
Synthasil reagent
Manufactured by Werfen
Sourced in Germany
SynthASil reagent is a laboratory product designed for use in analytical processes. It serves as a core component in various testing and analysis procedures conducted in scientific and medical laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Acl top 700cts, by Werfen (1 mentions)
Hemosil liquid antithrombin, by Werfen (1 mentions)
Hemosil protein c, by Werfen (1 mentions)
Hemosil free protein s, by Werfen (1 mentions)
Hemosil fibrinogen c xl reagent, by Werfen (1 mentions)
Acl top, by Werfen (1 mentions)
Recombiplastin 2g, by Werfen (1 mentions)
Acl top 700 system, by Werfen (1 mentions)
4 protocols using synthasil reagent
1
EHL-rFVIII Calibration Curve Development
2
Coagulation Assay Protocol: ACL TOP 700 CTS
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The aPTT and PT were performed on an ACL TOP 700 CTS (Werfen), and CWA was done using the integrated software. The aPTT was performed using the SynthasIL reagent (Werfen), and the PT was performed with the ReadiPlasTin reagent (Werfen). Plasma samples were mixed with the aPTT reagent, and the reaction was triggered with the addition of a solution of 20 mM CaCl2, or they were mixed with the PT reagent, according to the manufacturer's recommendations.
For the PT, the software was set up by the manufacturer to provide the time to Max1 (considered as the PT clotting time by the instrument and reported in seconds) and the Max1 (delta milliabsorbance [dmAbs]/dt). For aPTT, the software was set up to provide the Max1 (dmAbs/dt), the Max2 (dmAbs/dt2), the time to Max2 (considered as the aPTT clotting time by the instrument and reported in seconds) and the Min2 (dmAbs/dt2) (FigureS1 ).
For the PT, the software was set up by the manufacturer to provide the time to Max1 (considered as the PT clotting time by the instrument and reported in seconds) and the Max1 (delta milliabsorbance [dmAbs]/dt). For aPTT, the software was set up to provide the Max1 (dmAbs/dt), the Max2 (dmAbs/dt2), the time to Max2 (considered as the aPTT clotting time by the instrument and reported in seconds) and the Min2 (dmAbs/dt2) (Figure
3
Comprehensive Coagulation Profile Analysis
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Coagulation tests, including PT, aPTT, and factor assays, were performed on an automated coagulation analyzer, ACL TOP (Instrumentation Laboratory, Lexington, MA, USA). PT was measured using the HemosIL RecombiPlasTin reagent (Instrumentation Laboratory,), and aPTT was measured using the SynthASil reagent (Instrumentation Laboratory). Fibrinogen was measured using the HemosIL Fibrinogen-C XL reagent (Instrumentation Laboratory) based on the Clauss method. Coagulation factors were tested using a PT-based clotting assay with the HemosIL RecombiPlasTin reagent (for FV) and an aPTT-based clotting assay using the SynthASil reagent (for FVIII, FXI, and FXII). Antithrombin and protein C activity were determined with chromogenic assays (HemosIL liquid antithrombin and HemosIL Protein C; Instrumentation Laboratory) and free protein S antigen with an immunoassay (HemosIL Free Protein S, Instrumentation Laboratory).
4
Automated Coagulation Measurements
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The coagulation measurements were performed automatically on an ACL TOP 700 system from Instrumentation Laboratory (Kirchheim, Germany). For the prothrombin time, we used RecombiPlasTin 2G and for partial thromboplastin time SynthASil reagent, both from Instrumentation Laboratory (Kirchheim, Germany). All laboratory tests were performed automatically at the control temperature of 37°C and the clotting formation was measured by a turbidimetric method for dabigatran determination and the color development was measured at 405 nm for rivaroxaban determination, respectively. For calibration and control commercially available calibrators for dabigatran and rivaroxaban, as well as quality-control samples for both drugs from Technoclone GmbH (Vienna, Austria) were used.
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