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Heparin is not required for detection of antibodies associated with heparin-induced thrombocytopenia/thrombosis

  • Gian Paolo Visentin
    Affiliations
    Blood Research Institute, The Blood Center of Southeastern Wisconsin; Genetic Testing Institute Inc; and the Departments of Medicine and Pathology, Medical College of Wisconsin. Milwaukee, Wisconsin
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  • Michael Moghaddam
    Affiliations
    Blood Research Institute, The Blood Center of Southeastern Wisconsin; Genetic Testing Institute Inc; and the Departments of Medicine and Pathology, Medical College of Wisconsin. Milwaukee, Wisconsin
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  • Sheila E. Beery
    Affiliations
    Blood Research Institute, The Blood Center of Southeastern Wisconsin; Genetic Testing Institute Inc; and the Departments of Medicine and Pathology, Medical College of Wisconsin. Milwaukee, Wisconsin
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  • Janice G. McFarland
    Affiliations
    Blood Research Institute, The Blood Center of Southeastern Wisconsin; Genetic Testing Institute Inc; and the Departments of Medicine and Pathology, Medical College of Wisconsin. Milwaukee, Wisconsin
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  • Richard H. Aster
    Affiliations
    Blood Research Institute, The Blood Center of Southeastern Wisconsin; Genetic Testing Institute Inc; and the Departments of Medicine and Pathology, Medical College of Wisconsin. Milwaukee, Wisconsin
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      Abstract

      Heparin-induced thrombocytopenia (HIT), with or without thrombosis, is a common and often serious complication of heparin therapy. Platelet-activating, heparin-induced antibodies characteristic of HIT are thought to be specific for complexes formed between platelet factor 4 (PF4) and heparin, and such complexes are routinely used for antibody detection. We studied the binding of HIT antibodies to PF4 complexed with heparin fractions of uniform molecular size or linear polyanions other than heparin and found that many compounds other than heparin form complexes with PF4 that are suitable for antibody detection, provided they carry strong negative charges spaced about 0.5 nm apart along the molecular backbone and are of sufficient length to span about 40% of the circumference of the PF4 tetramer. Polyvinyl phosphonate was among the compounds that were equivalent to heparin. Thus neither a polysaccharide chain nor sulfate side groups—the hallmarks of heparin structure—are required for HIT antibody detection. The findings support the view that antibodies associated with HIT are specific for conformational changes that take place in the positively charged PF4 molecule when it reacts with a suitable, linear polyanion. (J Lab Clin Med 2001;138:22-31)

      Abbreviations:

      ELISA (enzyme-linked immunospecific assay), GAG (glycosaminoglycan), HF (heparin fragment), HIT (heparin-induced thrombocytopenia), IgG (immunoglobulin G), LMWH (low-molecular-weight heparin), OD (optical density), PAS (polyanethyl sulfonate), PBS (phosphate-buffered saline solution), PBS-Tween (PBS containing 0.05% Tween-20), PEG (polyethylene glycol), PF4 (platelet factor 4), PG (poly-D -glutamic acid), PSS (polystyrene sulfonate), PVP (polyvinyl phosphate), PVPN (polyvinyl phosphonate), PVS (polyvinyl sulfate), PVSN (polyvinyl sulfonate), SRA (serotonin release assay), UFH (unfractionated heparin)
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