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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   *
1   CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France
2   Laboratoire d’Hématologie Hôpital Universitaire Carémeau, Nîmes, France
3   Laboratoire d’Hématologie Hôpital Universitaire Saint-Eloi, Montpellier, France
,
Catherine Tarrade   *
1   CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France
,
Priscilla Lapalud
1   CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France
,
Sami Chtourou
4   LFB Biotechnologies, ZA Courtabeuf, les Ulis, France
,
Jean-François Schved
3   Laboratoire d’Hématologie Hôpital Universitaire Saint-Eloi, Montpellier, France
,
Claude Granier
1   CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France
,
Sylvie Villard-Saussine
1   CNRS FRE 3009, Cap Delta/Parc Euromédecine – Montpellier, France
› Author Affiliations
Further Information

Publication History

Received 07 August 2008

Accepted after major revision 20 April 2008

Publication Date:
27 November 2017 (online)

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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

* These authors contributed equally to this work.


 

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