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Inhibition of the procarboxypeptidase U (proCPU, TAFI, proCPB2) system due to hemolysis
Author(s): ,
Joachim C. Mertens
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
,
Karen Claesen
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
,
Dorien Leenaerts
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
,
Yani Sim
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
,
Anne‐Marie Lambeir
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
Dirk Hendriks
Affiliations:
Laboratory of Medical Biochemistry, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
Correspondence |Dirk Hendriks, Laboratory of Medical Biochemistry, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium.|E‐mail: dirk.hendriks@uantwerpen.be
ISTH Academy. Yadav R.
May 31, 2019; 273619
Rajesh Yadav
Rajesh Yadav
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Introduction
Spurious hemolysis of samples is the leading cause of interference in coagulation testing and was described to interfere in fibrinolysis assays. The influence of hemolysis on the procarboxypeptidase U (proCPU) system is not known.
Methods
By means of spiking of hemolysate in pooled normal plasma, the effect of hemolysis on CPU, proCPU, and functional clot lysis assays was assessed. The influence of hemolysis on CPU generation during in vitro clot lysis was also evaluated. Cutoffs corresponding to maximal acceptable bias were determined.
Results and discussion
When active CPU was added to pooled plasma, a severe decrease in activity – up to 97.2% inhibition – was seen with increasing plasma concentrations of oxyhemoglobin (oxyHb) and the 10% cutoff value was found to be 0.3 g/L oxyHb. Using an activity‐based assay, proCPU levels appeared to decrease gradually with increased hemolysis (maximal reduction of 19.5%) with a 10% cutoff value of 4.2 g/L oxyHb. The relative clot lysis time (CLT) showed a maximal negative bias of 68.5%. The reduction in CLT paralleled a significant reduction of the first CPU activity peak during clot lysis. The cutoff value for the CLT was 0.4 g/L oxyHb. In presence of thrombomodulin (TM), CLT+TM was not affected up to 8.0 g/L oxyHb.
Conclusion
These data indicate a clear inhibition of the CPU system because of hemolysis resulting in an increase of lysis in functional fibrinolysis assays. We were able to quantify the inhibitory effect and to propose cutoff values for every parameter.
Keyword(s)
carboxypeptidase B2, carboxypeptidase U, fibrinolysis, hemolysis, thrombin‐activatable fibrinolysis inhibitor
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