Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits

被引：0
作者：
Peter Chrenek
Lubos Ryban
Helga Vetr
Alexander V. Makarevich
Pavel Uhrin
Rekha K. Paleyanda
Bernd R. Binder
机构：
[1] Slovak Agricultural Research Centre,Department of Vascular Biology and Thrombosis Research
[2] Medical University of Vienna,undefined
[3] Technoclone GmbH,undefined
[4] American Integrated Biologics,undefined
[5] Inc.,undefined
[6] Massachusetts General Hospital,undefined
来源：
Transgenic Research | 2007年 / 16卷
关键词：
Recombinant; Clotting factor VIII; Mammary gland; Expression; Transgenic rabbit;
D O I：
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学科分类号：
摘要：
Transgenic founder rabbits carrying a gene construct consisting of a 2.5 kb murine whey acidic protein promoter (mWAP), 7.2 kb of the human clotting factor VIII (hFVIII) cDNA and 4.6 kb of 3′ flanking sequences of mWAP gene were crossed for three generations. All transgenic animals showed stable transgene transmission. Transgenic females showed high level of recombinant hFVIII (rhFVIII) mRNA expression in biopsed mammary gland tissues, while marginal expression of rhFVIII mRNA was observed in the spleen, lung and brain. No adverse effects of ectopic expression on the physiology of the rabbits were observed. Expression was not detected in the liver, kidney, heart and skeletal muscle. In transgenic females derived from three generations, rhFVIII protein was secreted from the mammary gland of lactating females, as shown by Western blotting. Biological activity of rhFVIII protein, as revealed in clotting assays was ranged from 0.012 to 0.599 IU/ml corresponding to 1.2% and 59.9% of the hFVIII level in normal human plasma. No apparent effect of secreted rhFVIII on the milk performance of rabbits was observed. Our results confirm the possibility of producing a significant amount of a biologically active rhFVIII in the mammary gland of established transgenic rabbit lines.
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页码：353 / 361
页数：8
相关论文
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