التفاصيل البيبلوغرافية
| العنوان: |
Factor XIIIa generation assay: A tool for studying factor XIII function in plasma. |
| المؤلفون: |
Dodt, Johannes1 johannes.dodt@pei.de, Volkers, Peter1, Seitz, Rainer1 |
| المصدر: |
Analytical Biochemistry. Aug2013, Vol. 439 Issue 2, p145-151. 7p. |
| مصطلحات موضوعية: |
*BLOOD coagulation factor XIIIa, *BLOOD plasma, *BIOLOGY experiments, *THROMBIN, *CHEMICAL sample preparation, *DOSE-response relationship in biochemistry |
| مستخلص: |
Abstract: Triggering the extrinsic coagulation pathway in plasma and using a fluorogenic factor XIIIa (FXIIIa) substrate for continuously monitoring FXIIIa activity, an FXIIIa generation curve is obtained. The parameters area under the curve (AUC), time to peak (TTP), and concentration at peak (CP) were calculated. In dilutions of normal plasma in FXIII-deficient plasma, AUC and CP showed linear dose–response relationships, whereas TTP increased from 9.9min for 25% FXIII to 11.6min for 100% FXIII. Three FXIII-A preparations (rFXIII, rFXIII(V34L), and cellular FXIII [cFXIII]) showed a linear dose response for AUC and CP. The TTP increased slightly for rFXIII from 13.5 to 15.0min, but surprisingly for cFXIII TTP increased concentration dependently from 13.5 to 28.7min. Adding 5μg/ml FXIII-B at a concentration of 1U of FXIII-A increased the AUC for rFXIII(V34L) and cFXIII by approximately 20% and accelerated TTP from 27.3 to 20.8min for cFVIII, indicating a supportive function of FXIII-B in orientating cFXIII-A for thrombin cleavage. A commercial assay quantifying FXIII after complete activation in a restricted time window did not reveal differences in the cFXIII preparation with or without FXIII-B. The FXIIIa generation assay provides additional information about activation and function of FXIII. This advantage was underlined in experiments with an irreversible FXIIIa inhibitor. [Copyright &y& Elsevier] |
| قاعدة البيانات: |
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