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Optimization of the thrombin generation test components to measure potency of factor VIII concentrates.
Haemophilia. 2016 Sep; 22(5):780-9.H

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. 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.

Authors+Show Affiliations

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.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA.Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, USA. mikhail.ovanesov@fda.hhs.gov.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27038076

Citation

Jha, N K., et al. "Optimization of the Thrombin Generation Test Components to Measure Potency of Factor VIII Concentrates." Haemophilia : the Official Journal of the World Federation of Hemophilia, vol. 22, no. 5, 2016, pp. 780-9.
Jha NK, Shestopal SA, Gourley MJ, et al. Optimization of the thrombin generation test components to measure potency of factor VIII concentrates. Haemophilia. 2016;22(5):780-9.
Jha, N. K., Shestopal, S. A., Gourley, M. J., Woodle, S. A., Liang, Y., Sarafanov, A. G., Weinstein, M., & Ovanesov, M. V. (2016). Optimization of the thrombin generation test components to measure potency of factor VIII concentrates. Haemophilia : the Official Journal of the World Federation of Hemophilia, 22(5), 780-9. https://doi.org/10.1111/hae.12943
Jha NK, et al. Optimization of the Thrombin Generation Test Components to Measure Potency of Factor VIII Concentrates. Haemophilia. 2016;22(5):780-9. PubMed PMID: 27038076.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimization of the thrombin generation test components to measure potency of factor VIII concentrates. AU - Jha,N K, AU - Shestopal,S A, AU - Gourley,M J, AU - Woodle,S A, AU - Liang,Y, AU - Sarafanov,A G, AU - Weinstein,M, AU - Ovanesov,M V, Y1 - 2016/04/01/ PY - 2016/02/16/accepted PY - 2016/4/3/entrez PY - 2016/4/3/pubmed PY - 2017/2/24/medline KW - factor VIII KW - haemophilia KW - potency KW - thrombin generation test SP - 780 EP - 9 JF - Haemophilia : the official journal of the World Federation of Hemophilia JO - Haemophilia VL - 22 IS - 5 N2 - 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. SN - 1365-2516 UR - https://www.unboundmedicine.com/medline/citation/27038076/Optimization_of_the_thrombin_generation_test_components_to_measure_potency_of_factor_VIII_concentrates_ L2 - https://doi.org/10.1111/hae.12943 DB - PRIME DP - Unbound Medicine ER -