Transgenic pigs produce functional human factor VIII in milk

@article{Paleyanda1997TransgenicPP,
  title={Transgenic pigs produce functional human factor VIII in milk},
  author={R. Paleyanda and W. Velander and Timothy K. Lee and D. Scandella and F. Gwazdauskas and J. Knight and L. Hoyer and W. Drohan and H. Luboń},
  journal={Nature Biotechnology},
  year={1997},
  volume={15},
  pages={971-975}
}
Deficiency or abnormality of coagulation factor VIII (FVIII) causes a bleeding disorder called hemophilia A. Treatment involves FVIII concentrates prepared from pooled human plasma or recombinant FVIII (rFVIII) prepared from mammalian cell culture. The cost of highly purified FVIII or rFVIII is a major factor in hemophilia therapy and restricts prophylaxis. We have sought to generate a new source of rFVIII by targeting expression of the human FVIII cDNA to the mammary gland of transgenic pigs… Expand
Expression of factor VIII in recombinant and transgenic systems.
TLDR
The contemporary methods used for FVIII expression in mammalian cell culture systems are reviewed and the factors responsible for insufficient recoveries of rFVIII are discussed, and design of FV III with prolonged half-life in vivo is considered as another promising direction in improving rF VIII protein and efficiency of hemophilia A therapy. Expand
Temporal and spatial expression of biologically active human factor VIII in the milk of transgenic mice driven by mammary-specific bovine α-lactalbumin regulation sequences
TLDR
A hybrid gene containing bovine α-lactalbumin and human FVIII cDNA was constructed for microinjection into the pronuclei of newly fertilized mouse eggs and confirmed to be successfully integrated and stably germ-line transmitted. Expand
Functional factor VIII made with von Willebrand factor at high levels in transgenic milk
TLDR
226/N6 was secreted into milk as a biologically active single‐chain molecule that retained high specific activity, similar to therapeutic‐grade FVIII, and provides proof‐of‐principle for the study of expression of 226/N 6 and perhaps other single-chain bioengineered rFVIIIs in the milk of transgenic livestock. Expand
Enhanced recombinant human blood coagulation factor VIII expression in the milk of goats using replication-defective adenovirus
TLDR
A simple, rapid, and low-cost protein production method based on a replication-defective adenoviral vector system for recombinant human blood coagulation factor VIII and other biopharmaceutical interest is reported. Expand
Transgenic Rabbits as a Model Organism for Production of Human Clotting Factor VIII
TLDR
The generation of transgenic rabbits is reported as a model species for testing the expression of hFVIII in the mammary gland, and the transgene was identified in one female founder animal, and it was transmitted to the offspring in a Mendelian fashion, thus demonstrating stable integration of the gene construct into the germline of the transgenic rabbit. Expand
Expression of active human blood clotting factor VIII in mammary gland of transgenic rabbits.
TLDR
Results indicate the suitability of the transgenic rabbit mammary gland for rhFVIII production and show germ-line transmission of a single integrated copy, and a homozygous line was established from this animal. Expand
Production of recombinant human von Willebrand factor in the milk of transgenic pigs.
TLDR
Evidence is provided that production of rhvWF from transgenic pig milk is a potentially valuable technology and can be used as a cost-effective alternative in clinical applications. Expand
Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits
TLDR
The results confirm the possibility of producing a significant amount of a biologically active rhFVIII in the mammary gland of established transgenic rabbit lines and show stable transgene transmission in transgenic animals. Expand
PRODUCTION OF RECOMBINANT HUMAN COAGULATION FACTOR IX BY TRANSGENIC PIG
The milk-borne plasmin, identical to the one in the plasma, is the predominant protease in the milk. Producing of recombinant human coagulation factor IX (tg-rhFIX) in milk by mammary gland ofExpand
Pilot production of recombinant human clotting factor IX from transgenic sow milk.
TLDR
The overall manufacturing process presented here has the potential for industrial production of rhFIX for treatment of hemophilia B patients and high-quality rhFIX product was produced. Expand
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References

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Expression of the blood-clotting factor-VIII cDNA is repressed by a transcriptional silencer located in its coding region.
TLDR
It is established that fVIII cDNA contains sequences that dominantly inhibit its own expression from retroviral as well as from plasmid vectors. Expand
Elimination of a major inhibitor epitope in factor VIII.
TLDR
An active, recombinant hybrid human/porcine fVIII molecule is constructed by replacing the putative human fV III A2 domain epitope with the homologous porcine sequence and retained full activity in the presence of antibodies with specificity restricted to the human A2 epitope. Expand
Expression of Human Anti-Hemophilic Factor IX in the Milk of Transgenic Sheep
TLDR
A hybrid gene to direct the synthesis of human anti-hemophilic factor IX to the mammary gland is designed and introduced into sheep, and two transgenic ewes, each carrying about 10 copies of the foreign gene, have been analysed for expression. Expand
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TLDR
The increasing acceptance of recombinant FVIII (rFVIII) by both clinicians and patients has provided added impetus to the search for new sources of plasma proteins, and the benefits and limitations of transgenic production of human blood proteins are discussed. Expand
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TLDR
Analysis of the transgenic protein indicated that it had a significantly different carbohydrate composition from the recombinant enzyme produced in mouse C127 cells, and molecular size analysis of the oligosaccharides confirmed their differences and showed that the mouse cell-derived preparation contained larger, complex-type N-linked oligOSaccharide structures than the material produced in goat mammary tissue. Expand
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TLDR
Seven monoclonal antibodies raised against purified VIII:C were used to construct a preliminary epitope map of these polypeptides, consistent with the published description of the complete primary structure of VII:C and its thrombin cleavage products. Expand
Effect of von Willebrand factor coexpression on the synthesis and secretion of factor VIII in Chinese hamster ovary cells.
TLDR
It is reported that coexpression of vWF with factor VIII in Chinese hamster ovary cells resulted in increased stable accumulation of factor VIII activity in the absence of serum in the growth medium, and it is demonstrated that in ChineseHamster Ovary cells both factor VIII mRNA accumulation and the processing and secretion of the primary factor VIII translation product are inefficient processes. Expand
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The human factor VIII procoagulant protein (VIIIC) was purified from the VIIIC--factor VIII--related antigen (VIIIRAg) complex in commercial factor VIII concentrate by immunoadsorbent chromatographyExpand
Sequences in the coding region of clotting factor VIII act as dominant inhibitors of RNA accumulation and protein production.
TLDR
Results indicate that expression of FVIII is regulated by signals within F VIII coding sequence that result in decreased RNA accumulation and FV III protein production. Expand
A Scalable Method for the Purification of Recombinant Human Protein C from the Milk of Transgenic Swine
TLDR
The results demonstrate that transgenic pigs can be efficient “bioreactors” for the production of rHPC, and that the purification of the protein from milk by conventional methods can be developed for economical manufacture. Expand
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