title: Engineering soya bean seeds as a scalable platform to produce cyanovirin-N, a non-ARV microbicide against HIV.
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.
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA.
authors.forename: Barry R
authors.initials: BR
authors.lastname: O'Keefe
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: André M
authors.initials: AM
authors.lastname: Murad
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: Giovanni R
authors.initials: GR
authors.lastname: Vianna
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA.
authors.forename: Koreen
authors.initials: K
authors.lastname: Ramessar
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA.|Leidos, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
authors.forename: Carrie J
authors.initials: CJ
authors.lastname: Saucedo
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Wilson
authors.affiliation: ImQuest BioSciences LLC., Frederick, MD, USA.
authors.forename: Karen W
authors.initials: KW
authors.lastname: Buckheit
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: Nicolau B
authors.initials: NB
authors.lastname: da Cunha
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: Ana Claudia G
authors.initials: AC
authors.lastname: Araújo
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: Cristiano C
authors.initials: CC
authors.lastname: Lacorte
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.|Division of Clinical Sciences, St. George's, University of London, London, UK.
authors.forename: Luisa
authors.initials: L
authors.lastname: Madeira
authors.affiliation: Molecular Targets Laboratory, Center for Cancer Research, National Cancer Institute, NIH, Frederick, MD, USA.
authors.forename: James B
authors.initials: JB
authors.lastname: McMahon
authors.affiliation: EMBRAPA Genetic Resources and Biotechnology, Laboratory of Synthetic Biology, Brasília, DF, Brazil.
authors.forename: Elibio L
authors.initials: EL
authors.lastname: Rech
mesh_terms: D019380:Anti-HIV Agents; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering; D012639:Seeds / Q000235:genetics* / Q000378:metabolism; D013025:Glycine max / Q000235:genetics* / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; C107733:cyanovirin N
keywords: Cyanovirin-N; anti-HIV; soya bean