title: Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging.
abstract: Responding to the need for recombinant acidic fibroblast growth factor in the pharmaceutical and cosmetic industries, we established a scalable expression system for recombinant human aFGF using transient and a DNA replicon vector expression inNicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered followingAgrobacteriuminfiltration ofN. benthamiana. The optimal time point at which to harvest recombinant human acidic fibroblast growth factor expressing leaves was found to be 4 days post-infiltration, before necrosis was evident. Commassie-stained SDS-PAGE gels of His-tag column eluates, concentrated using a 10 000 molecular weight cut-off column, showed an intense band at the expected molecular weight for recombinant human acidic fibroblast growth factor. An immunoblot confirmed that this band was recombinant human acidic fibroblast growth factor. Up to 10 µg recombinant human-acidic fibroblast growth factor/g of fresh leaves were achieved by a simple affinity purification protocol using protein extract from the leaves of agroinfiltratedN. benthamiana. The purified recombinant human acidic fibroblast growth factor improved the survival rate of UVB-irradiated HaCaT and CCD-986sk cells approximately 89 and 81 %, respectively.N. benthamiana-derived recombinant human acidic fibroblast growth factor showed similar effects on skin cell proliferation and UVB protection compared to those ofEscherichia coli-derived recombinant human acidic fibroblast growth factor. Additionally,N. benthamiana-derived recombinant human acidic fibroblast growth factor increased type 1 procollagen synthesis up to 30 % as well as reduced UVB-induced intracellular reactive oxygen species generation in fibroblast (CCD-986sk) cells.UVB is a well-known factor that causes various types of skin damage and premature aging. Therefore, the present study demonstrated thatN. benthamiana-derived recombinant human acidic fibroblast growth factor effectively protects skin cell from UVB, suggesting its potential use as a cosmetic or therapeutic agent against skin photoaging.
authors.affiliation: Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
authors.forename: Jang-Ho
authors.initials: JH
authors.lastname: Ha
authors.affiliation: Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea.
authors.forename: Ha-Neul
authors.initials: HN
authors.lastname: Kim
authors.affiliation: Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
authors.forename: Ki-Beom
authors.initials: KB
authors.lastname: Moon
authors.affiliation: Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
authors.forename: Jae-Heung
authors.initials: JH
authors.lastname: Jeon
authors.affiliation: Medical Nanomaterial Institute, Bio-FD&C, Hwasun, Jeonnam 58141, Korea.
authors.forename: Dai-Hyun
authors.initials: DH
authors.lastname: Jung
authors.affiliation: Medical Nanomaterial Institute, Bio-FD&C, Hwasun, Jeonnam 58141, Korea.
authors.forename: Su-Jung
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Biodesign Institute, School of Life Sciences, Arizona State University, Tempe, AZ, USA.
authors.forename: Hugh S
authors.initials: HS
authors.lastname: Mason
authors.affiliation: Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea.
authors.forename: Seo-Yeon
authors.initials: SY
authors.lastname: Shin
authors.affiliation: Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
authors.forename: Hyun-Soon
authors.initials: HS
authors.lastname: Kim
authors.affiliation: Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea.
authors.forename: Kyung-Mok
authors.initials: KM
authors.lastname: Park
mesh_terms: D060054:Agrobacterium; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000494:pharmacology* / Q000633:toxicity; D005822:Genetic Vectors; D006801:Humans; D030821:Plants, Genetically Modified; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D012867:Skin / Q000187:drug effects / Q000528:radiation effects; D015595:Skin Aging / Q000187:drug effects*; D014026:Nicotiana / Q000235:genetics*; D014466:Ultraviolet Rays
chemical_list: D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1