Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging

Jang Ho Ha, Ha Neul Kim, Ki Beom Moon, Jae Heung Jeon, Dai Hyun Jung, Su Jung Kim, Hugh Mason, Seo Yeon Shin, Hyun Soon Kim, Kyung Mok Park

Research output: Contribution to journalArticle

4 Citations (Scopus)

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 in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. 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 agroinfiltrated N. 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 of Escherichia 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 that N. 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.

Original languageEnglish (US)
JournalPlanta Medica
DOIs
StateAccepted/In press - Feb 7 2017

Fingerprint

Skin Aging
Fibroblast Growth Factor 1
Tobacco
Skin
Cosmetics
Charge coupled devices
Infiltration
Molecular Weight
Molecular weight
Premature Aging
Agrobacterium
Replicon
Cell proliferation
Drug Industry
Fibroblasts
Collagen Type I
Escherichia coli
Purification
Polyacrylamide Gel Electrophoresis
Reactive Oxygen Species

Keywords

  • DNA replicon vector
  • Nicotiana benthamiana
  • recombinant human aFGF
  • skin photoaging
  • Solanaceae
  • transient expression
  • UVB irradiation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Drug Discovery
  • Complementary and alternative medicine
  • Organic Chemistry

Cite this

Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging. / Ha, Jang Ho; Kim, Ha Neul; Moon, Ki Beom; Jeon, Jae Heung; Jung, Dai Hyun; Kim, Su Jung; Mason, Hugh; Shin, Seo Yeon; Kim, Hyun Soon; Park, Kyung Mok.

In: Planta Medica, 07.02.2017.

Research output: Contribution to journalArticle

Ha, Jang Ho ; Kim, Ha Neul ; Moon, Ki Beom ; Jeon, Jae Heung ; Jung, Dai Hyun ; Kim, Su Jung ; Mason, Hugh ; Shin, Seo Yeon ; Kim, Hyun Soon ; Park, Kyung Mok. / Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging. In: Planta Medica. 2017.
@article{5ef778aa98dd45f5886189350cbfb4d8,
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 in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. 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 agroinfiltrated N. 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 of Escherichia 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 that N. 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.",
keywords = "DNA replicon vector, Nicotiana benthamiana, recombinant human aFGF, skin photoaging, Solanaceae, transient expression, UVB irradiation",
author = "Ha, {Jang Ho} and Kim, {Ha Neul} and Moon, {Ki Beom} and Jeon, {Jae Heung} and Jung, {Dai Hyun} and Kim, {Su Jung} and Hugh Mason and Shin, {Seo Yeon} and Kim, {Hyun Soon} and Park, {Kyung Mok}",
year = "2017",
month = "2",
day = "7",
doi = "10.1055/s-0043-103964",
language = "English (US)",
journal = "Planta Medica",
issn = "0032-0943",
publisher = "Georg Thieme Verlag",

}

TY - JOUR

T1 - Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging

AU - Ha, Jang Ho

AU - Kim, Ha Neul

AU - Moon, Ki Beom

AU - Jeon, Jae Heung

AU - Jung, Dai Hyun

AU - Kim, Su Jung

AU - Mason, Hugh

AU - Shin, Seo Yeon

AU - Kim, Hyun Soon

AU - Park, Kyung Mok

PY - 2017/2/7

Y1 - 2017/2/7

N2 - 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 in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. 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 agroinfiltrated N. 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 of Escherichia 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 that N. 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.

AB - 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 in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. 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 agroinfiltrated N. 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 of Escherichia 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 that N. 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.

KW - DNA replicon vector

KW - Nicotiana benthamiana

KW - recombinant human aFGF

KW - skin photoaging

KW - Solanaceae

KW - transient expression

KW - UVB irradiation

UR - http://www.scopus.com/inward/record.url?scp=85014093308&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014093308&partnerID=8YFLogxK

U2 - 10.1055/s-0043-103964

DO - 10.1055/s-0043-103964

M3 - Article

C2 - 28249301

AN - SCOPUS:85014093308

JO - Planta Medica

JF - Planta Medica

SN - 0032-0943

ER -