Journal of thrombosis and haemostasis : JTH · 2026
Characterization of monoclonal and patient-derived antiplatelet factor 4 antibodies in platelet factor 4 and platelet factor 4/polyanion chemiluminescence assays.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Dabbiru Venkata A S, Broto Marta, Wesche Jan, Wang Jing Jing, Müller Luisa, Gebicka Patrycja, Rollin Jérôme, Gruel Yves, Palicio Marta, Thiele Thomas, Greinacher Andreas, Schönborn Linda
Аннотация
BACKGROUND: Heparin-induced thrombocytopenia, vaccine-induced immune thrombocytopenia and thrombosis (VITT), and VITT-like disorders are caused by antiplatelet factor 4 (PF4)/polyanion and/or anti-PF4 immunoglobulin G antibodies. The rapid anti-PF4/polyanion and anti-PF4 chemiluminescence assays differentiate between typical heparin-induced thrombocytopenia (anti-PF4/heparin) and VITT antibodies (anti-PF4).
OBJECTIVES: To characterize monoclonal antibodies (mAbs) and recombinant anti-PF4 antibodies in the 2 chemiluminescence assays specific for anti-PF4/polyanion and anti-PF4 antibodies, respectively, and to characterize anti-PF4/polyanion antibodies in patients whose sera tested positive in both assays.
METHODS: For anti-PF4/heparin and anti-PF4 antibodies, the chimeric mAbs 5B9, 1C12, and 1E12 were generated using standard technology. Recombinant antibodies from VITT and VITT-like patients were generated based on the amino acid sequences of affinity-purified patient anti-PF4 antibodies. KKO was humanized by cloning its Fab sequences and expressing them as a chimeric recombinant human immunoglobulin G1 antibody. Anti-PF4 antibodies from patient sera testing positive in both chemiluminescence assays were affinity-purified using PF4/polyanion and PF4 beads.
RESULTS: In chemiluminescence assays, 5B9, humanized KKO, 1C12, and 1E12 bound to PF4/polyanion complexes but differed largely in their PF4 reactivity. VITT (-like) recombinant antibodies bound only to PF4. Some patients' sera contained antibodies that showed similar reactivity in both assays, regardless of whether purified with PF4/polyanion or PF4 beads, suggesting cross-reactive antibodies, while sera from other patients contained both anti-PF4/heparin and anti-PF4 antibodies.
CONCLUSION: The different reactivities of anti-PF4 mAbs/heparin antibodies need to be considered when planning experiments and interpreting results. Patients may have very different antibody reactivities despite giving the same results in the 2 chemiluminescence assays.
OBJECTIVES: To characterize monoclonal antibodies (mAbs) and recombinant anti-PF4 antibodies in the 2 chemiluminescence assays specific for anti-PF4/polyanion and anti-PF4 antibodies, respectively, and to characterize anti-PF4/polyanion antibodies in patients whose sera tested positive in both assays.
METHODS: For anti-PF4/heparin and anti-PF4 antibodies, the chimeric mAbs 5B9, 1C12, and 1E12 were generated using standard technology. Recombinant antibodies from VITT and VITT-like patients were generated based on the amino acid sequences of affinity-purified patient anti-PF4 antibodies. KKO was humanized by cloning its Fab sequences and expressing them as a chimeric recombinant human immunoglobulin G1 antibody. Anti-PF4 antibodies from patient sera testing positive in both chemiluminescence assays were affinity-purified using PF4/polyanion and PF4 beads.
RESULTS: In chemiluminescence assays, 5B9, humanized KKO, 1C12, and 1E12 bound to PF4/polyanion complexes but differed largely in their PF4 reactivity. VITT (-like) recombinant antibodies bound only to PF4. Some patients' sera contained antibodies that showed similar reactivity in both assays, regardless of whether purified with PF4/polyanion or PF4 beads, suggesting cross-reactive antibodies, while sera from other patients contained both anti-PF4/heparin and anti-PF4 antibodies.
CONCLUSION: The different reactivities of anti-PF4 mAbs/heparin antibodies need to be considered when planning experiments and interpreting results. Patients may have very different antibody reactivities despite giving the same results in the 2 chemiluminescence assays.