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Thromb Haemost 1989; 61(02): 238-242
DOI: 10.1055/s-0038-1646566
Original Article
Schattauer GmbH Stuttgart

In Vitro Carboxylation of a Blood Coagulation Factor IX Precursor Produced by Recombinant-DNA Technology

Berry A M Soute‡
1   The Department of Biochemistry, University of Limburg, Maastricht, The Netherlands
,
Alain Balland‡
2   Transgene, Strasbourg, France
,
Thérèse Faure‡
2   Transgene, Strasbourg, France
,
Henri de la Salle‡
2   Transgene, Strasbourg, France
,
Cees Vermeer
1   The Department of Biochemistry, University of Limburg, Maastricht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 18 July 1988

Accepted after revision 14 October 1988

Publication Date:
30 June 2018 (online)

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Summary

Blood coagulation factor IX (Christmas factor) is a plasma protein which is required for normal haemostasis. A functional deficiency of factor IX results in haemophilia B, a bleeding disorder which is generally treated by infusions of factor IX concentrates prepared from pooled human plasma. The use of human blood products is connected with the risk of transmitting viral agents responsible for diseases such as hepatitis B and AIDS. Recombinant DNA techniques may provide the means to produce the required proteins without exposing the patients to these risks and at lower costs. One of the problems which has to be overcome before recombinant factor IX can be used for therapeutical purposes is related to the vitamin K-dependent carboxylation of its 12 NH2-terminal glutamate residues. In cell cultures this carboxylation, which is required to render the protein its procoagulant activity, is far from complete, especially at high expression levels. In this paper we describe the in vitro carboxylation of non and/or partly carboxylated recombinant factor IX produced by transformed Chinese hamster ovary cells. The identity of the newly formed Gla residues was verified and it could be demonstrated that all carboxyl groups had been incorporated into the recombinant factor IX.

Keywords

Vitamin K - Carboxylase - Haemophilia B - Factor IX
 

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