Optimization of the thrombin generation test components to measure potency of factor VIII concentrates
Abstract
Introduction The thrombin generation test (TGT) is used both as a global haemostasis assay, and to compare activities of coagulation factor concentrates that have been spiked into patient plasma. However, TGT has not been systematically optimized to evaluate factor VIII (FVIII) product potency. Aims To improve the sensitivity of TGT to FVIII and allow a comparative analysis of the thrombin generating capacities of FVIII concentrates against reference preparations with known FVIII activity. Methods Concentrations of TGT components (analytical variables) were assessed to maximize the linearity and range of responses to the concentration of FVIII. Results We optimized the range and sensitivity of the TGT assay with respect to FVIII through the addition of FXIa to the assay. Other parameters that were adjusted, i.e. tissue factor (TF), procoagulant lipids and plasma concentrations, did not improve the ability of the assay to measure both high and very low levels of FVIII. In the optimized TF/FXIa‐activated TGT assay, all thrombin generation curve parameters were suitable for FVIII quantification, but thrombin peak height and maximal velocity demonstrated better linearity in the desired FVIII range. We found that the optimized TF/FXIa‐activated TGT has a wider range of sensitivity to FVIII than a commercially available TGT.more »
- Authors:
-
[2]
- Office of Blood Research and Review Center for Biologics Evaluation and Research U.S. Food and Drug Administration Silver Spring MD USA, Department of Physics George Washington University Washington DC USA
- Office of Blood Research and Review Center for Biologics Evaluation and Research U.S. Food and Drug Administration Silver Spring MD USA
- Publication Date:
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1400745
- Resource Type:
- Publisher's Accepted Manuscript
- Journal Name:
- Haemophilia
- Additional Journal Information:
- Journal Name: Haemophilia Journal Volume: 22 Journal Issue: 5; Journal ID: ISSN 1351-8216
- Publisher:
- Wiley-Blackwell
- Country of Publication:
- United Kingdom
- Language:
- English
Citation Formats
Jha, N. K., Shestopal, S. A., Gourley, M. J., Woodle, S. A., Liang, Y., Sarafanov, A. G., Weinstein, M., and Ovanesov, M. V. Optimization of the thrombin generation test components to measure potency of factor VIII concentrates. United Kingdom: N. p., 2016.
Web. doi:10.1111/hae.12943.
Jha, N. K., Shestopal, S. A., Gourley, M. J., Woodle, S. A., Liang, Y., Sarafanov, A. G., Weinstein, M., & Ovanesov, M. V. Optimization of the thrombin generation test components to measure potency of factor VIII concentrates. United Kingdom. https://doi.org/10.1111/hae.12943
Jha, N. K., Shestopal, S. A., Gourley, M. J., Woodle, S. A., Liang, Y., Sarafanov, A. G., Weinstein, M., and Ovanesov, M. V. Fri .
"Optimization of the thrombin generation test components to measure potency of factor VIII concentrates". United Kingdom. https://doi.org/10.1111/hae.12943.
@article{osti_1400745,
title = {Optimization of the thrombin generation test components to measure potency of factor VIII concentrates},
author = {Jha, N. K. and Shestopal, S. A. and Gourley, M. J. and Woodle, S. A. and Liang, Y. and Sarafanov, A. G. and Weinstein, M. and Ovanesov, M. V.},
abstractNote = {Introduction The thrombin generation test (TGT) is used both as a global haemostasis assay, and to compare activities of coagulation factor concentrates that have been spiked into patient plasma. However, TGT has not been systematically optimized to evaluate factor VIII (FVIII) product potency. Aims To improve the sensitivity of TGT to FVIII and allow a comparative analysis of the thrombin generating capacities of FVIII concentrates against reference preparations with known FVIII activity. Methods Concentrations of TGT components (analytical variables) were assessed to maximize the linearity and range of responses to the concentration of FVIII. Results We optimized the range and sensitivity of the TGT assay with respect to FVIII through the addition of FXIa to the assay. Other parameters that were adjusted, i.e. tissue factor (TF), procoagulant lipids and plasma concentrations, did not improve the ability of the assay to measure both high and very low levels of FVIII. In the optimized TF/FXIa‐activated TGT assay, all thrombin generation curve parameters were suitable for FVIII quantification, but thrombin peak height and maximal velocity demonstrated better linearity in the desired FVIII range. We found that the optimized TF/FXIa‐activated TGT has a wider range of sensitivity to FVIII than a commercially available TGT. Additionally, we demonstrated that the TF/FXIa‐activated assay performs adequately by comparing potency measurements of five commercially available FVIII products using TGT and traditional chromogenic and one‐stage clotting assays. Conclusions The optimized TGT assay can be used to quantify and compare the thrombin generating capacities of FVIII concentrates.},
doi = {10.1111/hae.12943},
journal = {Haemophilia},
number = 5,
volume = 22,
place = {United Kingdom},
year = {Fri Apr 01 00:00:00 EDT 2016},
month = {Fri Apr 01 00:00:00 EDT 2016}
}
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Works referenced in this record:
A modified thrombin generation test for the measurement of factor VIII concentrates
journal, May 2003
- Mcintosh, J. H.; Owens, D.; Lee, C. A.
- Journal of Thrombosis and Haemostasis, Vol. 1, Issue 5
Coagulation procofactor activation by factor XIa: Factor VIII activation by factor Xia
journal, March 2010
- Whelihan, M. F.; Orfeo, T.; Gissel, M. T.
- Journal of Thrombosis and Haemostasis, Vol. 8, Issue 7
Major surgery in a severe haemophilia A patient with high titre inhibitor: use of the thrombin generation test in the therapeutic decision
journal, September 2005
- Dargaud, Y.; Lienhart, A.; Meunier, S.
- Haemophilia, Vol. 11, Issue 5
Preclinical efficacy and safety of rVIII-SingleChain (CSL627), a novel recombinant single-chain factor VIII
journal, August 2013
- Zollner, Sabine B.; Raquet, Elmar; Müller-Cohrs, Jochen
- Thrombosis Research, Vol. 132, Issue 2
Thrombin generation in vitro in the presence of by-passing agents in siblings with severe haemophilia A
journal, January 2010
- Klintman, J.; Berntorp, E.; Astermark, J.
- Haemophilia, Vol. 16, Issue 1
Potency determination of factor VIII and factor IX for new product labelling and postinfusion testing: challenges for caregivers and regulators
journal, January 2015
- Dodt, J.; Hubbard, A. R.; Wicks, S. J.
- Haemophilia, Vol. 21, Issue 4
Determining the impact of instrument variation and automated software algorithms on the TGT in hemophilia and normalized plasma
journal, September 2013
- Woodle, S. A.; Shibeko, A. M.; Lee, T. K.
- Thrombosis Research, Vol. 132, Issue 3
Correction of microplate location effects improves performance of the thrombin generation test
journal, January 2013
- Liang, Yideng; Woodle, Samuel A.; Shibeko, Alexey M.
- Thrombosis Journal, Vol. 11, Issue 1
Is some better than none: are TEG and TGA profiles different in severe FVIII-deficient patients with inhibitors?
journal, December 2014
- Salinas, V.; Carmona, R.; Mohammed, B. M.
- Haemophilia, Vol. 21, Issue 3
A Rebuttal: Issues with the Assay of Factor VIII Activity in Plasma and Factor VIII Concentrates
journal, January 2001
- Rosén, Steffen
- Thrombosis and Haemostasis, Vol. 86, Issue 10
Evaluation of the activated partial thromboplastin time assay for clinical monitoring of PEGylated recombinant factor VIII (BAY 94-9027) for haemophilia A
journal, January 2014
- Gu, J-M.; Ramsey, P.; Evans, V.
- Haemophilia, Vol. 20, Issue 4
Monitoring rFVIII prophylaxis dosing using global haemostasis assays
journal, March 2013
- AL Hawaj, M. A.; Martin, E. J.; Venitz, J.
- Haemophilia, Vol. 19, Issue 3
Calibrated automated thrombin generation and modified thromboelastometry in haemophilia A
journal, April 2009
- van Veen, Joost J.; Gatt, Alex; Bowyer, Annette E.
- Thrombosis Research, Vol. 123, Issue 6
Thrombin generation assay using factor IXa as a trigger to quantify accurately factor VIII levels in haemophilia A: CAT-based factor VIII assay and bleeding tendency
journal, August 2011
- Ninivaggi, M.; Dargaud, Y.; Van OERLE, R.
- Journal of Thrombosis and Haemostasis, Vol. 9, Issue 8
Diagnosis and management challenges in patients with mild haemophilia A and discrepant FVIII measurements
journal, February 2014
- Trossaert, M.; Lienhart, A.; Nougier, C.
- Haemophilia, Vol. 20, Issue 4
The effect of corn trypsin inhibitor and inhibiting antibodies for FXIa and FXIIa on coagulation of plasma and whole blood: comment
journal, July 2015
- Parunov, L. A.; Surov, S. S.; Tucker, E.
- Journal of Thrombosis and Haemostasis, Vol. 13, Issue 8
Feasibility of using thrombin generation assay (TGA) for monitoring of haemostasis during supplementation therapy in haemophilic patients without inhibitors
journal, May 2012
- Ay, Y.; Balkan, C.; Karapinar, D. Y.
- Haemophilia, Vol. 18, Issue 6
A Single Chain Variant of Factor VIII Fc Fusion Protein Retains Normal In Vivo Efficacy but Exhibits Altered In Vitro Activity
journal, November 2014
- Buyue, Yang; Liu, Tongyao; Kulman, John D.
- PLoS ONE, Vol. 9, Issue 11
A modified thrombin generation test for investigating very low levels of factor VIII activity in hemophilia A
journal, November 2009
- Matsumoto, Tomoko; Nogami, Keiji; Ogiwara, Kenichi
- International Journal of Hematology, Vol. 90, Issue 5
Functional characteristics of N8, a new recombinant FVIII: IN VITRO FUNCTION OF N8
journal, June 2010
- Christiansen, M. L. S.; Balling, K. W.; Persson, E.
- Haemophilia, Vol. 16, Issue 6
Thrombin Generation Testing for Monitoring Hemophilia Treatment: A Clinical Perspective
journal, October 2010
- Salvagno, Gian; Berntorp, Erik
- Seminars in Thrombosis and Hemostasis, Vol. 36, Issue 07
The use of the new ReFacto AF Laboratory Standard allows reliable measurement of FVIII:C levels in ReFacto AF mock plasma samples by a one-stage clotting assay: FVIII:C LEVELS IN PATIENTS RECEIVING REFACTO
journal, March 2011
- Pouplard, C.; Caron, C.; Aillaud, M. F.
- Haemophilia
Thrombomodulin-modified thrombin generation after in vivo recombinant factor VIII treatment in severe hemophilia A
journal, September 2008
- Dielis, A. W. J. H.; Balliel, W. M. R.; van Oerle, R.
- Haematologica, Vol. 93, Issue 9
Potency and mass of factor VIII in FVIII products
journal, January 2009
- Butenas, S.; Parhami-Seren, B.; Gissel, M. T.
- Haemophilia, Vol. 15, Issue 1
The impact of thrombin generation and rotation thromboelastometry on assessment of severity of factor XI deficiency
journal, August 2015
- Livnat, Tami; Shenkman, Boris; Martinowitz, Uri
- Thrombosis Research, Vol. 136, Issue 2
The technique of measuring thrombin generation with fluorogenic substrates: 3. The effects of sample dilution
journal, January 2009
- Smedt, Erik De; Wagenvoord, Rob; Hemker, Coen
- Thrombosis and Haemostasis, Vol. 101, Issue 01
Thrombin generation testing in routine clinical practice: are we there yet?
journal, September 2008
- van Veen, J. J.; Gatt, A.; Makris, M.
- British Journal of Haematology, Vol. 142, Issue 6
International comparative field study of N8 evaluating factor VIII assay performance: N8 FIELD STUDY EVALUATING FVIII ASSAYS
journal, March 2011
- Viuff, D.; Barrowcliffe, Tw.; Saugstrup, T.
- Haemophilia, Vol. 17, Issue 4
Evaluation of Thrombin Generation Assay in Patients With Hemophilia
journal, October 2014
- Haghpanah, Sezaneh; Bazrafshan, Asghar; Silavizadeh, Samir
- Clinical and Applied Thrombosis/Hemostasis, Vol. 22, Issue 4
Calibrated Automated Thrombin Generation Measurement in Clotting Plasma
journal, January 2003
- Hemker, H. Coenraad; Giesen, Peter; Al Dieri, Raed
- Pathophysiology of Haemostasis and Thrombosis, Vol. 33, Issue 1
Bleeding phenotype in carriers of haemophilia A does not correlate with thrombin generation
journal, November 2014
- Olsson, A.; Hellgren, M.; Berntorp, E.
- Haemophilia, Vol. 21, Issue 1
The Calibrated Automated Thrombogram (CAT): a universal routine test for hyper- and hypocoagulability
journal, January 2002
- Hemker, H. C.; Giesen, P.; AlDieri, R.
- Pathophysiology of Haemostasis and Thrombosis, Vol. 32, Issue 5-6
Clotting factor product administration and same-day occurrence of thrombotic events, as recorded in a large healthcare database during 2008-2013
journal, November 2015
- Ekezue, B. F.; Sridhar, G.; Ovanesov, M. V.
- Journal of Thrombosis and Haemostasis, Vol. 13, Issue 12
The Relative Importance of the Factors II, VII, IX and X for the Prothrombinase Activity in Plasma of Orally Anticoagulated Patients
journal, January 1989
- Xi, Ma; Béguin, S.; Hemker, H. C.
- Thrombosis and Haemostasis, Vol. 62, Issue 02
Comparative field study evaluating the activity of recombinant factor VIII Fc fusion protein in plasma samples at clinical haemostasis laboratories
journal, November 2013
- Sommer, J. M.; Moore, N.; McGuffie-Valentine, B.
- Haemophilia, Vol. 20, Issue 2
Thrombin generation assay using factor XIa to measure factors VIII and IX and their glycoPEGylated derivatives is robust and sensitive
journal, October 2015
- Waters, E. K.; Hilden, I.; Sørensen, B. B.
- Journal of Thrombosis and Haemostasis, Vol. 13, Issue 11
Studies on the Different Modes of Action of the Anticoagulant Protease Inhibitors DX-9065a and Argatroban: I. EFFECTS ON THROMBIN GENERATION
journal, October 2002
- Nagashima, Hajime
- Journal of Biological Chemistry, Vol. 277, Issue 52
Characterization of F8 defects in haemophilia A in Pakistan: investigation of correlation between mutation type and the in vitro thrombin generation assay
journal, October 2013
- Khanum, F.; Collins, P. W.; Harris, R. L.
- Haemophilia, Vol. 20, Issue 2
From Principle to Practice: Bridging the Gap in Patient Profiling
journal, January 2013
- Foley, Jonathan H.; Orfeo, Thomas; Undas, Anetta
- PLoS ONE, Vol. 8, Issue 1
Thrombin generation and whole blood viscoelastic assays in the management of hemophilia: current state of art and future perspectives
journal, March 2013
- Young, Guy; Sørensen, Benny; Dargaud, Yesim
- Blood, Vol. 121, Issue 11
Thrombin generation and phenotypic correlation in haemophilia A
journal, July 2005
- Beltran-Miranda, C. P.; Khan, A.; Jaloma-Cruz, A. R.
- Haemophilia, Vol. 11, Issue 4
The effect of tissue factor concentration on calibrated automated thrombography in the presence of inhibitor bypass agents
journal, April 2009
- van VEEN, J. J.; Gatt, A.; Bowyer, A. E.
- International Journal of Laboratory Hematology, Vol. 31, Issue 2
Heparin neutralization is essential for accurate measurement of factor VIII activity and inhibitor assays in blood samples drawn from implanted venous access devices
journal, July 2000
- Manco-Johnson, Marilyn J.; Nuss, Rachelle; Jacobson, Linda J.
- Journal of Laboratory and Clinical Medicine, Vol. 136, Issue 1