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

Roger L. Lundblad; Jennifer Bergstrom; Rene De Vreker; Gordon Bray; Edward Gomperts; Don Baker; Henry S. Kingdon; Kenneth G. Mann; Jeanette Hartshorn; Richard J. Jenny

doi:10.1055/s-0037-1615363

Thrombosis and Haemostasis, Table of Contents

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

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Jennifer Bergstrom

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Rene De Vreker

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Gordon Bray

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Edward Gomperts

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Don Baker

¹From the Hyland/Immuno Division of Baxter Healthcare, Duarte and Glendale, CA

,

Henry S. Kingdon

²Hyland/Immuno, Baxter Healthcare, Lake Cook, IL

,

Kenneth G. Mann

³Department of Biochemistry, University of Vermont, Burlington, VT

,

Jeanette Hartshorn

⁴Haematologic Technologies Inc., Essex Junction, VT, USA

,

Richard J. Jenny

³Department of Biochemistry, University of Vermont, Burlington, VT

› Author Affiliations

Abstract

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.

Full Text

References

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