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Thromb Haemost 1998; 80(05): 811-815
DOI: 10.1055/s-0037-1615363
Review Article
Schattauer GmbH

Measurement of Active Coagulation Factors in Autoplex®-T with Colorimetric Active Site-Specific Assay Technology

Roger L. Lundblad
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Jennifer Bergstrom
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Rene De Vreker
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Gordon Bray
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Edward Gomperts
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Don Baker
1   From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA
,
Henry S. Kingdon
2   Hyland/Immuno, Baxter Healthcare, Lake Cook, IL
,
Kenneth G. Mann
3   Department of Biochemistry, University of Vermont, Burlington, VT
,
Jeanette Hartshorn
4   Haematologic Technologies Inc., Essex Junction, VT, USA
,
Richard J. Jenny
3   Department of Biochemistry, University of Vermont, Burlington, VT
› Author Affiliations
Further Information

Publication History

Received 30 October 1997

Accepted after resubmission 08 July 1998

Publication Date:
07 December 2017 (online)

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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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