Epitope mapping of human factor VIII inhibitor antibodies by deletion analysis of factor VIII fragments expressed in Escherichia coli.
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- D Scandella
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
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- S DeGraaf Mahoney
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
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- M Mattingly
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
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- D Roeder
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
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- L Timmons
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
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- C A Fulcher
- Department of Molecular Biology Development, Rorer Biotechnology, King of Prussia, PA 19406.
抄録
<jats:p>Epitopes for antibodies that inhibit factor VIII procoagulant protein were analyzed by deletion mapping of factor VIII protein fragments expressed in Escherichia coli. A human factor VIII cDNA clone was used to generate E. coli expression vectors encoding fragments containing the 80-kDa factor VIII light chain (A3, C1, and C2 domains) and the 44-kDa carboxyl-terminal half of the factor VIII heavy chain (A2 domain). A series of deletions of each fragment was constructed and tested by immunoblotting for the binding of alloantibody and autoantibody inhibitors. Analysis of derivatives of the 80-kDa fragment showed that six inhibitors recognized a major epitope(s) within the carboxyl-terminal 17.3 kDa of factor VIII. These inhibitors also recognized weaker epitopes nearby and one inhibitor recognized epitopes scattered throughout the 80-kDa fragment. Deletions within the heavy chain fragment revealed one epitope-containing region confined to the amino-terminal 18.3 kDa recognized by six inhibitors. Bacterially produced factor VIII fragments containing the major epitopes were capable of neutralizing inhibitors in vitro but fragments containing weaker or no epitopes did not. These data suggest a potential therapeutic use of factor VIII fragments for neutralization of inhibitor antibodies.</jats:p>
収録刊行物
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 85 (16), 6152-6156, 1988-08
Proceedings of the National Academy of Sciences
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詳細情報
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- CRID
- 1363107370918199296
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- NII論文ID
- 30016283488
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- ISSN
- 10916490
- 00278424
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- データソース種別
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