Combinatorial peptides directed to inhibitory antibodies against human blood clotting factor VIII

Eva-Maria Kopecky; Sabine Greinstetter; Ingrid Pabinger; Andrea Buchacher; Jürgen Römisch; Alois Jungbauer

doi:10.1160/TH05-04-0254

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 94(05): 933-941
DOI: 10.1160/TH05-04-0254

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Combinatorial peptides directed to inhibitory antibodies against human blood clotting factor VIII

Eva-Maria Kopecky

¹Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

,

Sabine Greinstetter

¹Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

,

Ingrid Pabinger

²Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University Vienna, Austria

,

Andrea Buchacher

³Octapharma Pharmazeutika Produktions Ges. mbH, Vienna, Austria

,

Jürgen Römisch

³Octapharma Pharmazeutika Produktions Ges. mbH, Vienna, Austria

,

Alois Jungbauer

¹Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

› Author Affiliations

Abstract

Summary

The development of antibodies against blood clotting factorVIII is a major complication affecting 20–30% of hemophilia A patients receiving replacement with FVIII concentrates. This study investigated generating peptides acting as broadly neutralizing agents to block factor VIII antibodies. These peptides were selected from dual positional scanning decapeptide libraries on cellulose membranes. From this library comprising 6.8×10¹² peptides we selected 468 peptides for further screening rounds. Finally we identified two decapeptides with the ability to block 8 out of 10 inhibitory antibodies from sera of patients with FVIII inhibitors demonstrated by competition assays. Sequence alignment of the peptides showed similarity with several domains in the FVIII molecule demonstrating the mimotope nature of the selected peptides. Our results show the efficiency of the combinatorial library approach and show the potential of combinatorial peptides to compete out polyclonal inhibitor IgG from a broad range of patients’ sera. Combinatorial peptides could be novel and highly effective drug candidates for alternative treatment in patients with factor VIII inhibitors.

Keywords

Hemophilia - factor VIII inhibitor - mimotopes - combinatorial peptides

Full Text

References

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