title: Adeno-associated-virus-mediated transduction of the mammary gland enables sustained production of recombinant proteins in milk.
abstract: Biopharming for the production of recombinant pharmaceutical proteins in the mammary gland of transgenic animals is an attractive but laborious alternative compared to mammalian cell fermentation. The disadvantage of the lengthy process of genetically modifying an entire animal could be circumvented with somatic transduction of only the mammary epithelium with recombinant, replication-defective viruses. While other viral vectors offer very limited scope for this approach, vectors based on adeno-associated virus (AAV) appear to be ideal candidates because AAV is helper-dependent, does not induce a strong immune response and has no association with disease. Here, we sought to test the suitability of recombinant AAV (rAAV) for biopharming. Using reporter genes, we showed that injected rAAV efficiently transduced mouse mammary cells. When rAAV encoding human myelin basic protein (hMBP) was injected into the mammary glands of mice and rabbits, this resulted in the expression of readily detectable protein levels of up to 0.5 g/L in the milk. Furthermore we demonstrated that production of hMBP persisted over extended periods and that protein expression could be renewed in a subsequent lactation by re-injection of rAAV into a previously injected mouse gland.
authors.affiliation: AgResearch, Ruakura, Hamilton, New Zealand.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wagner
authors.affiliation: AgResearch, Ruakura, Hamilton, New Zealand.
authors.forename: Rosemary
authors.initials: R
authors.lastname: Thresher
authors.affiliation: AgResearch, Grasslands, Palmerston North, New Zealand.
authors.forename: Ross
authors.initials: R
authors.lastname: Bland
authors.affiliation: AgResearch, Ruakura, Hamilton, New Zealand.
authors.forename: Götz
authors.initials: G
authors.lastname: Laible
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D042361:Mammary Glands, Human / Q000502:physiology*; D051379:Mice; D008894:Milk Proteins / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D008894:Milk Proteins; D011994:Recombinant Proteins