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DOI: 10.1055/s-0037-1615363
Measurement of Active Coagulation Factors in Autoplex®-T with Colorimetric Active Site-Specific Assay Technology
Publication History
Received
30 October 1997
Accepted after resubmission
08 July 1998
Publication Date:
07 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199805/lookinside/thumbnails/10-1055-s-0037-1615363-1.jpg)
Summary
Autoplex® -T is a partially activated prothrombin complex concentrate used primarily for the treatment of patients expressing factor VIII inhibitors. While Autoplex® -T has a demonstrated record of clinical effectiveness, the procoagulant composition of this material has not been reported. This absence of composition data is a reflection of the lack of techniques appropriate for accurately measuring an individual protease such as factor IXa in complex mixtures of similar proteases. The development of Colorimetric Active Site-Specific ImmunoAssay technology (CASSIA) has permitted the accurate analysis of the coagulant enzymes present in Autoplex® -T. Ten lots of Autoplex® -T were reacted with both biotinylated phenylalanylprolylarginine chloromethylketone and biotinylated glutamylglycylarginine chloromethylketone. Only activated forms of the clotting factors present in Autoplex® -T react with the peptide chloromethylketones and were thus separated from the other proteins present in Autoplex® -T by adsorption onto streptavidin. The individual proteins bound to streptavidin were then detected with specific antibodies. Mean results from the analysis of ten lots of Autoplex® -T (mean values) are as follows: factor XIa, 5.9 nM or 0.8 μg/ml; factor Xa, 46.5 nM or 2.1 μg/ml; factor IXa, 177.8 nM or 11.7 μg/ml; factor VIIa, 68.6 nM or 3.3 μg/ml and factor IIa, 5.3 nM or 0.2 μg/ml. These results are discussed with respect to the mechanism of action of Autoplex® -T in the treatment of factor VIII inhibitor patients.
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