Development and characterization of recombinant ovine coagulation factor VIII.
Development and characterization of recombinant ovine coagulation factor VIII.
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DOI：
10.1371/journal.pone.0049481
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Doering CB
中科院分区：
文献类型：
--
作者：
Zakas PM;Gangadharan B;Almeida-Porada G;Porada CD;Spencer HT;Doering CB
Animal models of the bleeding disorder, hemophilia A, have been an integral component of the biopharmaceutical development process and have facilitated the development of recombinant coagulation factor VIII (fVIII) products capable of restoring median survival of persons with hemophilia A to that of the general population. However, there remain several limitations to recombinant fVIII as a biotherapeutic, including invasiveness of intravenous infusion, short half-life, immunogenicity, and lack of availability to the majority of the world's population. The recently described ovine model of hemophilia A is the largest and most accurate phenocopy. Affected sheep die prematurely due to bleeding-related pathogenesis and display robust adaptive humoral immunity to non-ovine fVIII. Herein, we describe the development and characterization of recombinant ovine fVIII (ofVIII) to support further the utility of the ovine hemophilia A model. Full-length and B-domain deleted (BDD) ofVIII cDNAs were generated and demonstrated to facilitate greater biosynthetic rates than their human fVIII counterparts while both BDD constructs showed greater expression rates than the same-species full-length versions. A top recombinant BDD ofVIII producing baby hamster kidney clone was identified and used to biosynthesize raw material for purification and biochemical characterization. Highly purified recombinant BDD ofVIII preparations possess a specific activity nearly 2-fold higher than recombinant BDD human fVIII and display a differential glycosylation pattern. However, binding to the carrier protein, von Willebrand factor, which is critical for stability of fVIII in circulation, is indistinguishable. Decay of thrombin-activated ofVIIIa is 2-fold slower than human fVIII indicating greater intrinsic stability. Furthermore, intravenous administration of ofVIII effectively reverses the bleeding phenotype in the murine model of hemophilia A. Recombinant ofVIII should facilitate the maintenance of the ovine hemophilia A herd and their utilization as a relevant large animal model for the research and development of novel nucleic acid and protein-based therapies for hemophilia A.
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DOI：
10.1111/j.1538-7836.2009.03697.x
发表时间：
2010-02
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Porada CD;Sanada C;Long CR;Wood JA;Desai J;Frederick N;Millsap L;Bormann C;Menges SL;Hanna C;Flores-Foxworth G;Shin T;Westhusin ME;Liu W;Glimp H;Zanjani ED;Lozier JN;Pliska V;Stranzinger G;Joerg H;Kraemer DC;Almeida-Porada G
通讯作者：
Almeida-Porada G
影响因子：
12.4
作者：
Doering, Christopher B.;Denning, Gabriela;Spencer, H. Trent
通讯作者：
Spencer, H. Trent
影响因子：
4.8
作者：
Doering, CB;Healey, JF;Lollar, P
通讯作者：
Lollar, P
影响因子：
2
作者：
Bihoreau, N;Veillon, JF;Schmitter, JM
通讯作者：
Schmitter, JM
影响因子：
4.2
作者：
Dooriss, Kerry L.;Denning, Gabriela;Doering, Christopher B.
通讯作者：
Doering, Christopher B.