alexa Whole blood thrombin: development of a process for intra-operative production of human thrombin.
Pharmaceutical Sciences

Pharmaceutical Sciences

Biochemistry & Pharmacology: Open Access

Author(s): Kumar V, Chapman JR

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Abstract Thrombin-based clotting agents currently used for topical hemostasis with absorbable sponges, fibrin sealants, and platelet gels are primarily derived from bovine or pooled human plasma sources. Autologous thrombin has important safety advantages in that it does not carry the same safety concerns as pooled plasma-derived products and it avoids exposure to risks associated with bovine-derived proteins. The goal of our research was to develop a rapid, reliable, and simple to perform process to generate autologous human thrombin in the intraoperative setting, from patient whole blood as the starting source material. Using whole blood instead of plasma as the starting material, it is possible to avoid the inherent delay in thrombin availability associated with a primary step of plasma isolation. In this study, we varied several key processing parameters to maximize thrombin production, reproducibility and stability. Autologous thrombin production was generated using a dedicated, single use disposable with a sterile reagent. The disposable consists of a tubular reaction chamber containing glass microsphere beads to activate the alternative pathway of the coagulation cascade. At the end of the process, thrombin-activated serum was harvested from the reaction chamber. The average activity of the thrombin produced at room temperature by this system was 82.8 +/- 15.9 IU/mL. The total processing time was < 30 minutes. The system was compatible with Anticoagulant Citrate Dextrose-Solution A (ACD-A) (8\%-12\%). The average volume of thrombin harvested from each aliquot of blood was 7.0 +/- 0.3 mL, and the stability of thrombin was observed to be temperature dependent, with cold storage better preserving thrombin activity. Clot times with platelet concentrates at 1:4.3 and 1:11 ratios (thrombin to platelet concentrate) were < 10 and 20 seconds, respectively. A process for the preparation of thrombin from whole blood, under conditions compatible with the resources of an operating room, has been developed. The device is simple to use, requires 30 minutes, and can consistently produce thrombin solutions that achieve rapid clotting of platelet concentrates, plasma, and fibrinogen concentrates even when mixed at thrombin to blood product ratios of 1:11.
This article was published in J Extra Corpor Technol and referenced in Biochemistry & Pharmacology: Open Access

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