Characterization of anticoagulant heparinoids by immunoprofiling |
| |
Authors: | Tessa J Wijnhoven Els M van de Westerlo Nicole C Smits Joost F Lensen Angelique L Rops Johan van der Vlag Jo H Berden Lambert P van den Heuvel Toin H van Kuppevelt |
| |
Institution: | (1) Department of Matrix Biochemistry, Nijmegen Centre for Molecular Life Sciences, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands;(2) Department of Pediatric Nephrology, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands;(3) Nephrology Research Laboratory, Division of Nephrology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands |
| |
Abstract: | Heparinoids are used in the clinic as anticoagulants. A specific pentasaccharide in heparinoids activates antithrombin III,
resulting in inactivation of factor Xa and–when additional saccharides are present–inactivation of factor IIa. Structural
and functional analysis of the heterogeneous heparinoids generally requires advanced equipment, is time consuming, and needs
(extensive) sample preparation. In this study, a novel and fast method for the characterization of heparinoids is introduced
based on reactivity with nine unique anti-heparin antibodies. Eight heparinoids were biochemically analyzed by electrophoresis
and their reactivity with domain-specific anti-heparin antibodies was established by ELISA. Each heparinoid displayed a distinct
immunoprofile matching its structural characteristics. The immunoprofile could also be linked to biological characteristics,
such as the anti-Xa/anti-IIa ratio, which was reflected by reactivity of the heparinoids with antibodies HS4C3 (indicative
for 3-O-sulfates) and HS4E4 (indicative for domains allowing anti-factor IIa activity). In addition, the immunoprofile could be indicative
for heparinoid-induced side-effects, such as heparin-induced thrombocytopenia, as illustrated by reactivity with antibody
NS4F5, which defines a very high sulfated domain. In conclusion, immunoprofiling provides a novel, fast, and simple methodology
for the characterization of heparinoids, and allows high-throughput screening of (new) heparinoids for defined structural
and biological characteristics. |
| |
Keywords: | Antibody Anticoagulation Heparin-induced thrombocytopenia Heparinoid Immunoprofiling |
本文献已被 PubMed SpringerLink 等数据库收录! |
|