Molecular characterization of human B domain-specific anti-factor VIII monoclonal antibodies generated in transgenic mice

Géraldine Lavigne-Lissalde; Sébastien Lacroix-Desmazes; Bharath Wootla; Catherine Tarrade; Jean-François Schved; Srini V. Kaveri; Claude Granier; Sylvie Villard-Saussine

doi:10.1160/TH06-09-0510

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2007; 98(01): 138-147
DOI: 10.1160/TH06-09-0510

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Molecular characterization of human B domain-specific anti-factor VIII monoclonal antibodies generated in transgenic mice

Géraldine Lavigne-Lissalde^*

¹1CNRS UMR 5160, Centre de Pharmacologie et Biotechnologie pour la Santé, Faculté de Pharmacie, Montpellier, France

²Groupe Hospitalo-Universitaire CHU Carémeau, Nîmes, France

,

Sébastien Lacroix-Desmazes^*

³Institut National de la Santé et de la Recherche Médicale (INSERM), Unité (U) 681, Paris, France

⁴Université Pierre et Marie Curie (UPMC – Paris 6), Centre de recherche des Cordeliers, Institut Fédératif de Recherche (IFR) 58, Paris, France

,

Bharath Wootla

³Institut National de la Santé et de la Recherche Médicale (INSERM), Unité (U) 681, Paris, France

⁴Université Pierre et Marie Curie (UPMC – Paris 6), Centre de recherche des Cordeliers, Institut Fédératif de Recherche (IFR) 58, Paris, France

,

Catherine Tarrade

¹1CNRS UMR 5160, Centre de Pharmacologie et Biotechnologie pour la Santé, Faculté de Pharmacie, Montpellier, France

,

Jean-François Schved

⁵Hôpital Universitaire Saint-Eloi, Montpellier, France

,

Srini V. Kaveri

³Institut National de la Santé et de la Recherche Médicale (INSERM), Unité (U) 681, Paris, France

⁴Université Pierre et Marie Curie (UPMC – Paris 6), Centre de recherche des Cordeliers, Institut Fédératif de Recherche (IFR) 58, Paris, France

,

Claude Granier

¹1CNRS UMR 5160, Centre de Pharmacologie et Biotechnologie pour la Santé, Faculté de Pharmacie, Montpellier, France

,

Sylvie Villard-Saussine

¹1CNRS UMR 5160, Centre de Pharmacologie et Biotechnologie pour la Santé, Faculté de Pharmacie, Montpellier, France

› Author Affiliations

Abstract

Summary

The development of antibodies directed against factorVIII (FVIII) represents a major hurdle in the treatment of hemophilia A. Most anti-FVIII antibodies are identified through their ability to inhibit the FVIII procoagulant activity. Many of them, however, do not interfere with the functional properties of FVIII. Antibodies directed against the B domain belong to this latter category. Here, we characterized B domain-specific human monoclonal Abs (mAbs) at the molecular level. A series of human mAbs directed against FVIII was produced upon immunization of transgenic XenoMouse mice with human recombinant FVIII (rFVIII). Selection of the hybridoma with epitope specificity for the B domain was performed by differential recognition of full-length and B domain-deleted rFVIII. None of the anti-B domain mAbs demonstrated inhibitory activity against FVIII. Three of the mAbs recognized linear epitopes: mAb 25H3 bound to the ¹⁰¹⁴HIDGPSLLIEN¹⁰²⁴ sequence; mAbs 8E3 and 22B6 shared the same epitope, composed of residues ¹⁵³⁴KWNEANR¹⁵⁴⁰. The corresponding soluble peptides inhibited the binding of their respective mAbs to FVIII. mAbs 8E3 and 22B6 displaced the binding of FVIII to vonWillebrand factor. Moreover, some of them (in particular mAbs 4G6 and 8E3) were able to compete for binding to the B domain with the anti-FVIII Abs from hemophilia A patients without inhibitor or with low Bethesda titers. Further investigation will allow to better characterize their clinical relevance.

Keywords

HemophiliaA - FVIII - anti-FVIII antibodies - XenoMouse - B domain

Full Text

References

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