SEARCH:   Advanced

Blood, 12 March 2009, Vol. 113, No. 11, pp. 2587-2594. Prepublished online as a Blood First Edition Paper on January 14, 2009; DOI 10.1182/blood-2008-08-174987. This Article Full Text Full Text (PDF) All Versions of this Article: blood-2008-08-174987v1 113/11/2587    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Krudysz-Amblo, J. Articles by Mann, K. G. Search for Related Content PubMed PubMed Citation Articles by Krudysz-Amblo, J. Articles by Mann, K. G. Related Collections Immunobiology Free Research Articles Thrombosis and Hemostasis Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  THROMBOSIS AND HEMOSTASIS Quantitation of anti–factor VIII antibodies in human plasma Jolanta Krudysz-Amblo1, Behnaz Parhami-Seren1, Saulius Butenas1, Kathleen E. Brummel-Ziedins1, Edward D. Gomperts2, Georges E. Rivard3, and Kenneth G. Mann1 1 Department of Biochemistry, College of Medicine, University of Vermont, Burlington; 2 Children's Hospital Los Angeles, CA; and 3 Centre Hospitalier Universitaire (CHU) Sainte-Justine, Montreal, QC The presence of antibodies (Abs) in hemophilia A patients can potentially influence the therapeutic qualities of factor VIII (fVIII) administration. Much work has been focused on the presence of inhibitory antibodies, whereas the quantitation of noninhibitory anti-fVIII antibodies has been largely undetermined. Our objective was to develop a sensitive and specific fluorescence-based immunoassay (FLI) for the quantitation of anti-fVIIIAbs in human plasma. Affinity-purified human anti-fVIIIAb, isolated from a hemophilia A subject, was used as a calibrator with a detectability limit of 40 (±1.5) pM. The calibrator and the human plasma anti-fVIIIAb were captured on recombinant fVIII (rfVIII)– coupled microspheres and probed with mouse anti–human Ig–R-phycoerythrin. Plasma samples from 150 healthy donors and 39 inhibitor-negative hemophilia A subjects were compared with 4 inhibitor-positive hemophilia A plasma samples with inhibitor titers of 1 BU/mL (94.6 ± 0.8 nM), 11 BU/mL (214.3 ± 7.1 nM), 106 BU/mL (2209.4 ± 84.9 nM), 140 BU/mL (2417.7 ± 3.8 nM) as measured by the Nijmegen method. We also describe the validation of a mouse anti–human fVIIIAb as a surrogate calibrator. Four healthy individuals (3%) showed detectable anti-fVIIIAb in the range of 0.6 to 6.2 nM, whereas 13 (33%) of the 39 inhibitor-free hemophilia A subjects were positive for anti-fVIIIAb in the range of 0.5 to 20 nM. The method may be useful for therapeutic management of hemophilia A patients. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

	Copyright © 2009 by American Society of Hematology         Online ISSN: 1528-0020