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Engineering soya bean seeds as a scalable platform to produce cyanovirin‐N,a non‐ARV microbicide against HIV
Authors:Barry R O'Keefe  André M Murad  Giovanni R Vianna  Koreen Ramessar  Carrie J Saucedo  Jennifer Wilson  Karen W Buckheit  Nicolau B da Cunha  Ana Claudia G Araújo  Cristiano C Lacorte  Luisa Madeira  James B McMahon  Elibio L Rech
Institution:1. Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA;2. EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil;3. Leidos, Frederick National Laboratory for Cancer Research, Frederick, MD, USA;4. ImQuest BioSciences LLC., Frederick, MD, USA;5. Division of Clinical Sciences, St. George's, University of London, London, UK
Abstract:There is an urgent need to provide effective anti‐HIV microbicides to resource‐poor areas worldwide. Some of the most promising microbicide candidates are biotherapeutics targeting viral entry. To provide biotherapeutics to poorer areas, it is vital to reduce the cost. Here, we report the production of biologically active recombinant cyanovirin‐N (rCV‐N), an antiviral protein, in genetically engineered soya bean seeds. Pure, biologically active rCV‐N was isolated with a yield of 350 μg/g of dry seed weight. The observed amino acid sequence of rCV‐N matched the expected sequence of native CV‐N, as did the mass of rCV‐N (11 009 Da). Purified rCV‐N from soya is active in anti‐HIV assays with an EC50 of 0.82–2.7 nM (compared to 0.45–1.8 nM for E. coli‐produced CV‐N). Standard industrial processing of soya bean seeds to harvest soya bean oil does not diminish the antiviral activity of recovered rCV‐N, allowing the use of industrial soya bean processing to generate both soya bean oil and a recombinant protein for anti‐HIV microbicide development.
Keywords:Cyanovirin‐N  soya bean  anti‐HIV
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