Simultaneous detection and epitope mapping of anti-factor VIII antibodies

Geraldine Lavigne-Lissalde; Catherine Tarrade; Priscilla Lapalud; Sami Chtourou; Jean-François Schved; Claude Granier; Sylvie Villard-Saussine

doi:10.1160/TH07-08-0497

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2008; 99(06): 1090-1096
DOI: 10.1160/TH07-08-0497

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

Simultaneous detection and epitope mapping of anti-factor VIII antibodies

Geraldine Lavigne-Lissalde^*

¹CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France

²Laboratoire d’Hématologie Hôpital Universitaire Carémeau, Nîmes, France

³Laboratoire d’Hématologie Hôpital Universitaire Saint-Eloi, Montpellier, France

,

Catherine Tarrade^*

¹CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France

,

Priscilla Lapalud

¹CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France

,

Sami Chtourou

⁴LFB Biotechnologies, ZA Courtabeuf, les Ulis, France

,

Jean-François Schved

³Laboratoire d’Hématologie Hôpital Universitaire Saint-Eloi, Montpellier, France

,

Claude Granier

¹CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France

,

Sylvie Villard-Saussine

¹CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France

› Author Affiliations

Abstract

Summary

The development of antibodies (Abs) against infused factor VIII (FVIII) is currently one of the most serious complications in the treatment of patients suffering from haemophilia A. Improved prevention and eradication of these anti-FVIII Abs remain a challenge for both clinicians and scientists. Here we describe an immunoassay to simultaneously detect and map the epitope specificity of haemophilia A patients’ inhibitors by screening plasma against both heavy and light chains (HC and LC) of human plasma-derived FVIII (pFVIII). The format used was a two-site sandwich assay, where one monoclonal antibody (mAb) specific for the HC or LC was first immobilized on beads, and then incubated with the different forms of pFVIII. After incubation with patients’ plasma samples, binding was revealed by a phycoerythrin-labeled secondary Ab. Samples from haemophilia patients with autoantibodies (autoAb) or alloantibodies (alloAb) were screened in this format. The former preferentially recognized the LC, whereas the latter were directed against both LC and HC. This technology appears attractive as it is fast and requires only 100 μl of patient’s plasma. Furthermore, not only are anti-FVIII Abs detected, but information on their epitopic specificity is also obtained.

Keywords

Haemophilia A - inhibitors - epitope mapping

Full Text

References

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