{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T03:31:08Z","timestamp":1790047868681,"version":"4.0.1"},"reference-count":20,"publisher":"Georg Thieme Verlag KG","issue":"08","license":[{"start":{"date-parts":[[2023,5,10]],"date-time":"2023-05-10T00:00:00Z","timestamp":1683676800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"CSL Behring Innovation GmbH"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Thromb Haemost"],"published-print":{"date-parts":[[2023,8]]},"abstract":"<jats:p>\n            Background\u2003There is an emerging concept that in addition to circulating coagulation factor IX (FIX), extravascular FIX contributes to hemostasis.<\/jats:p><jats:p>\n            Objective\u2003Our objective was to evaluate the efficacy of extravascular FIX using animal models of tail clip bleeding and ferric chloride-induced thrombosis.<\/jats:p><jats:p>\n            Methods\u2003Mutant rFIX proteins with described enhanced (rFIXK5R) or reduced (rFIXK5A) binding to extracellular matrix were generated and characterized using in vitro aPTT, one-stage clotting, and modified FX assays. Using hemophilia B mice, pharmacokinetic (PK) parameters and in vivo efficacy of these proteins were compared against rFIX wild-type protein (rFIXWT) in a tail clip bleeding and FeCl3-induced thrombosis model. Respective tissue disposition of FIX was evaluated using immunofluorescence.<\/jats:p><jats:p>\n            Results\u2003In vitro characterization demonstrated comparable clotting activity of rFIX proteins. The PK profile showed that rFIXK5A displayed the highest plasma exposure compared to rFIXWT and rFIXK5R. Immunofluorescence evaluation of liver tissue showed that rFIXK5R was detectable up to 24\u2009hours, whereas rFIXWT and rFIXK5A were detectable only up to 15\u2009minutes. In the tail clip bleeding model, rFIXK5R displayed significant hemostatic protection against bleeding incidence for up to 72\u2009hours postintravenous administration of 50\u2009IU\/kg, whereas the efficacy of rFIXK5A was already reduced at 24\u2009hours. Similarly, in the mesenteric artery thrombus model, rFIXK5R and rFIXWT demonstrated prolonged efficacy compared to rFIXK5A.<\/jats:p><jats:p>\n            Conclusion\u2003Using two different in vivo models of hemostasis and thrombosis, we demonstrate that mutated rFIX protein with enhanced binding (rFIXK5R) to extravascular space confers prolonged hemostatic efficacy in vivo despite lower plasma exposure, whereas rFIXK5A rapidly lost its efficacy despite higher plasma exposure.<\/jats:p>","DOI":"10.1055\/a-2090-9739","type":"journal-article","created":{"date-parts":[[2023,5,10]],"date-time":"2023-05-10T23:28:14Z","timestamp":1683761294000},"page":"751-762","source":"Crossref","is-referenced-by-count":9,"title":["Modulation of Extravascular Binding of Recombinant Factor IX Impacts the Duration of Efficacy in Mouse Models"],"prefix":"10.1055","volume":"123","author":[{"given":"Sabine","family":"Pestel","sequence":"first","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6941-8796","authenticated-orcid":false,"given":"Padmapriya","family":"Ponnuswamy","sequence":"first","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steffi Knoll","family":"Machado","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hendrik","family":"Peil","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timo","family":"Kraushaar","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philipp","family":"Claar","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel","family":"Mischnik","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holger","family":"Lind","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Herzog","sequence":"additional","affiliation":[{"name":"CSL Behring LLC, King of Prussia, Pennsylvania, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Bacher","sequence":"additional","affiliation":[{"name":"Institute of Immunology\u2014Philipps University Marburg, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc Wolfgang","family":"Nolte","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maximilian","family":"Bielohuby","sequence":"additional","affiliation":[{"name":"CSL Behring Innovation GmbH, Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2023,6,8]]},"reference":[{"issue":"03","key":"ref2","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1055\/s-0037-1615621","article-title":"Definitions in hemophilia. 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