TY - JOUR
T1 - Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF)
AU - He, Yonghua
AU - Schmidt, Monica A.
AU - Erwin, Christopher
AU - Guo, Jun
AU - Sun, Raphael
AU - Pendarvis, Ken
AU - Warner, Brad W.
AU - Herman, Eliot M.Herman
N1 - Funding Information:
This research was supported by NIH# R21 DK094065 (BWW, PI; EMH and MAS coPIs), University Research Strategic Alliance Program; Washington University in St. Louis, and the Children's Surgical Sciences Research Institute - St. Louis Children's Hospital Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
© 2016 He et al.
PY - 2016/6
Y1 - 2016/6
N2 - Necrotizing enterocolitis (NEC) is a devastating condition of premature infants that results from the gut microbiome invading immature intestinal tissues. This results in a life-threatening disease that is frequently treated with the surgical removal of diseased and dead tissues. Epidermal growth factor (EGF), typically found in bodily fluids, such as amniotic fluid, salvia and mother's breast milk, is an intestinotrophic growth factor and may reduce the onset of NEC in premature infants. We have produced human EGF in soybean seeds to levels biologically relevant and demonstrated its comparable activity to commercially available EGF. Transgenic soybean seeds expressing a seed-specific codon optimized gene encoding of the human EGF protein with an added ER signal tag at the N' terminal were produced. Seven independent lines were grown to homozygous and found to accumulate a range of 6.7 +/- 3.1 to 129.0 +/- 36.7 μg EGF/g of dry soybean seed. Proteomic and immunoblot analysis indicates that the inserted EGF is the same as the human EGF protein. Phosphorylation and immunohistochemical assays on the EGF receptor in HeLa cells indicate the EGF protein produced in soybean seed is bioactive and comparable to commercially available human EGF. This work demonstrates the feasibility of using soybean seeds as a biofactory to produce therapeutic agents in a soymilk delivery platform.
AB - Necrotizing enterocolitis (NEC) is a devastating condition of premature infants that results from the gut microbiome invading immature intestinal tissues. This results in a life-threatening disease that is frequently treated with the surgical removal of diseased and dead tissues. Epidermal growth factor (EGF), typically found in bodily fluids, such as amniotic fluid, salvia and mother's breast milk, is an intestinotrophic growth factor and may reduce the onset of NEC in premature infants. We have produced human EGF in soybean seeds to levels biologically relevant and demonstrated its comparable activity to commercially available EGF. Transgenic soybean seeds expressing a seed-specific codon optimized gene encoding of the human EGF protein with an added ER signal tag at the N' terminal were produced. Seven independent lines were grown to homozygous and found to accumulate a range of 6.7 +/- 3.1 to 129.0 +/- 36.7 μg EGF/g of dry soybean seed. Proteomic and immunoblot analysis indicates that the inserted EGF is the same as the human EGF protein. Phosphorylation and immunohistochemical assays on the EGF receptor in HeLa cells indicate the EGF protein produced in soybean seed is bioactive and comparable to commercially available human EGF. This work demonstrates the feasibility of using soybean seeds as a biofactory to produce therapeutic agents in a soymilk delivery platform.
UR - http://www.scopus.com/inward/record.url?scp=84976293894&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0157034
DO - 10.1371/journal.pone.0157034
M3 - Article
C2 - 27314851
AN - SCOPUS:84976293894
SN - 1932-6203
VL - 11
JO - PLoS ONE
JF - PLoS ONE
IS - 6
M1 - e0157034
ER -