African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12269

Full Length Research Paper

Enhanced recombinant human blood coagulation factor VIII expression in the milk of goats using replication-defective adenovirus

Zengsheng Han*
  • Zengsheng Han*
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Lin Wang
  • Lin Wang
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Kun Li
  • Kun Li
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Jian Li
  • Jian Li
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Hongwei Zhao
  • Hongwei Zhao
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Zhiwei Liu
  • Zhiwei Liu
  • Department of Biological Engineering, College of Environment and Chemical Engineering, Yanshan University, Qinhuangdao, Hebei Province 066004, China.
  • Google Scholar
Wenguang Cao
  • Wenguang Cao
  • Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China.
  • Google Scholar
Qingwang Li
  • Qingwang Li
  • College of Animal Science and Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi Province 712100, China.
  • Google Scholar


  •  Received: 11 February 2015
  •  Accepted: 22 April 2015
  •  Published: 29 April 2015

Abstract

High-level expression of recombinant human blood coagulation factor in milk of farm animals at a large scale provides a powerful tool for protein production. However, a bottleneck in recent protein synthesis technologies is the high cost of current transgenic livestock system. Here, we report a simple, rapid, and low-cost protein production method based on a replication-defective adenoviral vector system. The recombinant hfVIII adenoviral vector was generated by using homologous recombination in bacteria and transfected the plasmids into the HEK293 packaging cell line. Goat’s mammary glands at the different physiological stage were infected with the recombinant adenovirus containing a human blood coagulation factor VIII gene. The expression level of bioactive hfVIII from milk sample was confirmed by Western blot analysis and ELISA methods. The hfVIII gene was expressed as a protein of about 26 kDa and no recombinant hfVIII protein was detected in negative control treatments. The hfVIII was expressed with a high activity level up to 1.4 U/ml in the milk of goats. These data suggest that the approach established here could be a low cost and further increase expression efficacy for the recombinant human blood coagulation factor and other biopharmaceutical interest.
 
Key words: Adenovirus vector, human blood coagulation factor VIII, mammary gland, milk, goat, recombinant protein.