{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T04:59:43Z","timestamp":1761541183905,"version":"3.37.3"},"reference-count":36,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2014,4,21]],"date-time":"2014-04-21T00:00:00Z","timestamp":1398038400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/3.0\/"}],"funder":[{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"publisher","award":["10SDG3500034"],"award-info":[{"award-number":["10SDG3500034"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2014,9]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Factor <jats:styled-content style=\"fixed-case\">VIII<\/jats:styled-content> (<jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>) is a multidomain blood plasma glycoprotein. Activated <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> acts as a cofactor to the serine protease factor <jats:styled-content style=\"fixed-case\">IX<\/jats:styled-content>a within the membrane\u2010bound tenase complex assembled on the activated platelet surface. Defect or deficiency in <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> causes haemophilia A, a severe hereditary bleeding disorder. Intravenous administration of plasma\u2010derived <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> or recombinant <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> concentrates restores normal coagulation in haemophilia A patients and is used as an effective therapy. In this work, we studied the biophysical properties of clinically potent recombinant <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> forms: human <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> full\u2010length (<jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">FL<\/jats:styled-content>), human <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> B\u2010domain deleted (<jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">BDD<\/jats:styled-content>) and porcine <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">BDD<\/jats:styled-content> bound to negatively charged phospholipid vesicles at near\u2010physiological conditions. We used cryo\u2010electron microscopy (Cryo\u2010<jats:styled-content style=\"fixed-case\">EM<\/jats:styled-content>) as a direct method to evaluate the homogeneity and micro\u2010organization of the protein\u2010vesicle suspensions, which are important for <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> therapeutic properties. Applying concurrent Cryo\u2010<jats:styled-content style=\"fixed-case\">EM<\/jats:styled-content>, circular dichroism and dynamic light scattering studies to the three recombinant <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> forms when bound to phospholipid vesicles revealed novel properties for their functional, membrane\u2010bound state. The three <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> constructs have similar activity, secondary structure distribution and bind specifically to negatively charged phospholipid membranes. Human and porcine <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">BDD<\/jats:styled-content> induce strong aggregation of the vesicles, but the human <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">FL<\/jats:styled-content> form does not. The proposed methodology is effective in characterizing and identifying differences in therapeutic recombinant <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> membrane\u2010bound forms near physiological conditions, because protein\u2010containing aggregates are considered to be a factor in increasing the immunogenicity of protein therapeutics. This will provide better characterization and development of safer and more effective <jats:styled-content style=\"fixed-case\">FVIII<\/jats:styled-content> products with implications for haemophilia A treatment.<\/jats:p>","DOI":"10.1111\/hae.12421","type":"journal-article","created":{"date-parts":[[2014,4,21]],"date-time":"2014-04-21T06:20:53Z","timestamp":1398061253000},"page":"723-731","source":"Crossref","is-referenced-by-count":12,"title":["Lack of recombinant factor <scp>VIII<\/scp> B\u2010domain induces phospholipid vesicle aggregation: implications for the immunogenicity of factor <scp>VIII<\/scp>"],"prefix":"10.1111","volume":"20","author":[{"given":"K.","family":"Grushin","sequence":"first","affiliation":[{"name":"Department of Neuroscience and Cell Biology University of Texas Medical Branch Galveston TX USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J.","family":"Miller","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Cell Biology University of Texas Medical Branch Galveston TX USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"D.","family":"Dalm","sequence":"additional","affiliation":[{"name":"Department of Neuroscience and Cell Biology University of Texas Medical Branch Galveston TX USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E. 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