Sedimentation Velocity Analytical Ultracentrifugation of Oxidized Recombinant Full-Length Factor VIII
| العنوان: | Sedimentation Velocity Analytical Ultracentrifugation of Oxidized Recombinant Full-Length Factor VIII |
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| المؤلفون: | Pete Lollar, David Lillicrap, John F. Healey, Philip M. Zakas, Ian W. Smith |
| المصدر: | Frontiers in Immunology Frontiers in Immunology, Vol 11 (2020) |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | lcsh:Immunologic diseases. Allergy, 0301 basic medicine, Drug, congenital, hereditary, and neonatal diseases and abnormalities, oxidation, media_common.quotation_subject, animal diseases, Immunology, Excipient, Protein aggregation, immunogenicity, law.invention, 03 medical and health sciences, 0302 clinical medicine, Immune system, law, hemic and lymphatic diseases, medicine, Immunology and Allergy, Humans, media_common, Original Research, Factor VIII, biology, Chemistry, Immunogenicity, Recombinant Proteins, 3. Good health, 030104 developmental biology, Coagulation, Biochemistry, biology.protein, Recombinant DNA, hemophilia A, Antibody, lcsh:RC581-607, Oxidation-Reduction, Ultracentrifugation, analytical ultracentrifugation, 030215 immunology, medicine.drug |
| الوصف: | Anti-drug antibodies to coagulation factor VIII (fVIII), often termed inhibitors, present the greatest economical and treatment related obstacle in the management of hemophilia A. Although several genetic and environmental risk factors associated with inhibitor development have been identified, the precise mechanisms responsible for the immune response to exogenous fVIII therapies remain undefined. Clinical trials suggest there is an increased immunogenic potential of recombinant fVIII compared to plasma-derived products. Additional biochemical and immunological studies have demonstrated that changes in recombinant fVIII production and formulation can alter fVIII structure and immunogenicity. Recently, one study demonstrated increased immunogenicity of the recombinant fVIII product Helixate in hemophilia A mice following oxidation with hypochlorite (ClO−). It is widely reported that protein aggregates within drug products can induce adverse immune reactions in patients. Several studies have therefore investigated the prevalence of molecular aggregates in commercial recombinant products with and without use-relevant stress and agitation. To investigate the potential link between oxidation-induced immunogenicity and molecular aggregation, we analyzed the recombinant fVIII product, Helixate, via sedimentation velocity analytical ultracentrifugation following oxidation with ClO−. At 80 μM ClO−, a concentration that reduced the specific-activity by 67%, no detectable increase in large molecular aggregates (s > 12 S) was observed when compared to non-oxidized fVIII. This lack of aggregates was demonstrated both in commercial excipient as well as a HEPES buffered saline formulation. These data suggest that oxidation induced immunogenicity is independent of aggregate-mediated immune response. Therefore, our data support multiple, independent mechanisms underlying fVIII immunogenicity. |
| تدمد: | 1664-3224 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7578214e1c7636763f1b2cb76e6fbcc6 https://pubmed.ncbi.nlm.nih.gov/32117290 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....7578214e1c7636763f1b2cb76e6fbcc6 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16643224 |
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