Multiple VH genes are used to assemble human antibodies directed toward the A3-C1 domains of factor VIII

E. N. Van den Brink, E. A M Turenhout, N. Bovenschen, B. G A D H Heijnen, K. Mertens, M. Peters, J. Voorberg*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

A well-known complication of factor VIII replacement therapy in patients with hemophilia A is the development of inhibitory antibodies. Several studies have demonstrated the presence of a binding site for factor VIII inhibitors in the A3 domain. Six different human monoclonal single-chain variable domain antibody fragments (scFv) directed toward the A3-C1 domains of factor VIII have been isolated, using phage display technology. Sequence analysis revealed that the VH domains of 2 scFv were encoded by germline gene segments from the VH1 gene family and 4 by germline gene segments belonging to the VH3 gene family. Epitope mapping of the scFv was performed, using a series of hybrid factor VIII/factor V light chain fragments. This analysis revealed that 5 of 6 scFv were directed against a region encompassing amino acid sequence Q1778-D1840 in the A3 domain, a previously identified binding site for factor VIII inhibitors. Only 2 of 5 scFv directed against amino acid sequence Q1778-D1840 inhibited the procoagulant activity of factor VIII. Our results define the properties of human antibodies directed against region Q1778-D1840 in the A3 domain. Binding of one, noninhibitory scFv was independent of the region Q1778-D1840, suggesting the presence of an additional binding site for anti-factor VIII antibodies in the A3-C1 domains of factor VIII.

Original languageEnglish
Pages (from-to)966-972
Number of pages7
JournalBlood
Volume97
Issue number4
DOIs
Publication statusPublished - 15 Feb 2001
Externally publishedYes

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