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Thromb Haemost 1987; 58(04): 1043-1048
DOI: 10.1055/s-0038-1646053
Original Article
Schattauer GmbH Stuttgart

F VIII Subunits: Purification and Antigenic Properties

Ole Nordfang
Nordisk Gentofte AIS, Gentofte, Denmark
,
Mirella Ezban
Nordisk Gentofte AIS, Gentofte, Denmark
,
Jan J Hansen
Nordisk Gentofte AIS, Gentofte, Denmark
› Author Affiliations
Further Information

Publication History

Received 09 April 1987

Accepted after revision 24 August 1987

Publication Date:
29 June 2018 (online)

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Summary

Factor VIII-Light Chain (FVIII-LC) and FVIII-Heavy Chain (FVIII-HC) were purified from human plasma by the use of immunosorbents containing monoclonal antibodies or human inhibitor antibodies. FVIII-LC was subsequently isolated in essentially pure state by cation exchange chromatography. The preparations obtained contained 50 ng of protein for each unit of FVIII-LC antigen (FVIII-LC: Ag).

Affinity purified FVIII-LC and FVIII-HC preparations containing less than 0.3% of the opposite subunit were added in FVIILC inhibition assay of hemophilia A inhibitor antibodies. FVIII-LC was able to fully block the inhibitor activity in 6 out of 7 hemophilia A plasmas and partially block the inhibitor activity of one plasma. FVIII-HC only blocked FVIILC inhibiting antibodies form the plasma that was not fully blocked by FVIII-LC. It is suggested that FVIII-LC can be used for immunotherapy of the patients whose FVIILC inhibiting antibodies are directed towards FVIII-LC.

When FVIII-LC was coupled to Sepharose at a concentration of 4800 units of FVIII-LC: Ag per ml Sepharose, 0.2 ml of the immunosorbent was able to bind 900 Bethesda units from 100 ml hemophilia A inhibitor plasma. This opens the possibility to remove FVIII inhibitor antibodies from circulation by extracorporeal immunotherapy with FVIII-LC coupled to Sepharose.

Keywords

Factor VIII - Hemophilia A inhibitor antibodies - Factor VIII Light Chain - Factor VIII Heavy Chain
 

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