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Thromb Haemost 1987; 57(01): 102-105
DOI: 10.1055/s-0038-1651071
Original Articles
Schattauer GmbH Stuttgart

Immunoaffinity Purification of Factor VIII Complex

V S Hornsey
The Edinburgh and South-East Scotland Blood Transfusion Service, Royal Infirmary, University of Edinburgh Medical School, Edinburgh, Scotland
,
B D Griffin
*   The Scottish National Blood Transfusion Service Headquarters Unit Laboratory, University of Edinburgh Medical School, Edinburgh, Scotland
,
D S Pepper
*   The Scottish National Blood Transfusion Service Headquarters Unit Laboratory, University of Edinburgh Medical School, Edinburgh, Scotland
,
L R Micklem
**   The Department of Surgery, University of Edinburgh Medical School, Edinburgh, Scotland
,
C V Prowse
The Edinburgh and South-East Scotland Blood Transfusion Service, Royal Infirmary, University of Edinburgh Medical School, Edinburgh, Scotland
› Institutsangaben
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Publikationsverlauf

Received 25. August 1986

Accepted after revision 05. Dezember 1986

Publikationsdatum:
06. Juli 2018 (online)

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Summary

Murine monoclonal antibodies to human von Willebrand factor (vWf) were immobilised on Sephacryl S-1000. Various solutes were screened for their ability to elute 125 I-vWf from the immobilised antibodies. The most effective solutions were then tested to determine which allowed retention of factor VIII procoagulant activity (VIII: C) and activity of vWf measured by platelet aggregation in the presence of ristocetin (Ristocetin cofactor activity R.cof.). Finally, F VIII complex was purified from both plasma and cryoprecipitate by immunoaffinity chromatography under the selected conditions. The product had a specific activity of 45 units of VIII: C per mg of protein and 60 units of R. cof. per mg representing a 4000-fold purification from plasma. The fibrinogen and fibroneetin content were each less than 4% of the total protein with vWf accounting for 60% of the total protein in the final product. Multimer analysis of the product showed a similar pattern to normal plasma and contamination by murine monoclonal antibody was less than 300 ng per mg of protein. A novel product is thus obtained containing both clinically relevant VIII :C and R. cof. in a single vial whilst using only one specific monoclonal antibody.

Keywords

Factor VIII - Monoclonal antibody - Affinity chromatography
 

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