English

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.