Recombinant porcine EGF protein (Qk064)

Porcine EGF is an essential growth factor belonging to the epidermal growth factor family that stimulates the proliferation and differentiation of cells. It is used for the differentiation of induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) towards epithelial, neural, mesoderm and hematopoietic lineages.

Porcine EGF is becoming rapidly adopted as a species-specific growth factor in the cultivated meat sector. Porcine EGF is also an essential component of many organoid media supporting the culture and differentiation of a wide variety of organoids including intestine and liver organoids.

Porcine EGF has a molecular weight of 6.3 kDa. This protein is animal origin-free, carrier protein-free, His-tag-free and non-glycosylated to ensure a pure recombinant protein with exceptional lot-to-lot consistency.

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Summary

  • High purity porcine EGF protein (Uniprot: Q00968)

  • >98%, by SDS-PAGE quantitative densitometry

  • Expressed in E. coli
  • 6.3 kDa monomer
  • Animal origin-free (AOF)

  • Carrier protein-free
  • Manufactured in Cambridge, UK
  • Lyophilized from acetonitrile/TFA
  • Resuspend in 10mM HCl, prepare single-use aliquots, add carrier protein if desired, and store frozen at -20°C or -80°C

Featured applications

  • Differentiation of iPSCs into epithelial cells
  • Culture of porcine (pig) muscle stem cells
  • Culture of porcine (pig) organoids
  • Inclusion in cultivated meat media

Epidermal growth factor, EGF, pEGF, Porcine epidermal growth factor, Urogastrone, URG, HOMG4, beta-urogastrone, pro-epidermal growth factor, Epidermal growth factor (beta-urogastrone)

Porcine (pig)

Bioactivity

Qk064 pEGF_bioactivity_lot 204515

Porcine EGF (pEGF) and human EGF (hEGF) activity is determined using the Promega serum response element (SRE) luciferase reporter assay (*) in transfected HEK293T cells. Cells are treated in triplicate with a serial dilution of pEGF for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. pEGF (green) has an EC50 of 64 pg/ml (10.2 pM) which is comparable to hEGF (black) with an EC50 of 70.5 pg/mL (11.2 pM). Data from Qk064 lot #204515 and Qk011 lot #104419. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340

Purity

pEGF_purity graph (Qk064)

Recombinant pEGF migrates as a single band at approximately 13 kDa (monomer) in non-reducing (NR) conditions and an approximately 6 kDa band upon reduction. No contaminating protein bands are present. Purified recombinant protein (3 µg) was resolved using 18% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk064 lot #204515.

Further quality assays

  • Mass spectrometry, single species with the expected mass
  • Endotoxin: <0.005 EU/μg protein (below the level of detection)
  • Recovery from stock vial: >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Qkine porcine EGF and human EGF proteins have equivalent bioactivity

porcine EGF human EGF bioactivity comparison

Bioactivity was determined using a firefly luciferase reporter assay in stably transfected HEK293T cells. Cells were treated with a serial dilution of porcine EGF (Qk064, green) or human EGF (Qk011, black) for 3 hours in triplicate.

Protein background

Epidermal growth factor (EGF) is an essential potent growth factor that stimulates cell proliferation, differentiation, and survival of a variety of epidermal and epithelial cells. It is used extensively in induced-pluripotent stem cell (iPSC) and embryonic stem cell (ESC) culture systems for the successful expansion and differentiation of epithelial, neural, mesoderm and hematopoietic lineages [1-2]. It also plays a crucial role in wound healing [3].  EGF belongs to the EGF family comprising of TGF-alpha, amphiregulin (AR), betacellulin (BTC), epiregulin (EPR), heparin-binding EGF-like growth factor (HB-EGF), epigen and the neuregulins (NRGs) [4].

Porcine EGF is a globular protein, has a molecular weight of 6.3 kDa and composed of 53 amino acids. Structurally, it is a 3-dimensional protein consisting of a series of loops and three intramolecular disulfide bonds formed by six conserved cysteine residues. These bonds help to stabilise its structure and allow for receptor binding [5]. Generally, epidermal growth factor family members are synthesized as type I transmembrane precursor proteins, often containing several EGF domains in the extracellular region. The proteolytic cleavage of the EGF domain proximal to the transmembrane yields mature proteins that are released from the cell surface [4, 6].  EGF has been shown to be expressed in various tissues, organs and body fluids including blood, saliva, amniotic fluid and cerebrospinal fluid [7].

Epidermal growth factor binds to high-affinity EGF receptors (EGFRs) which belong to the family of receptor tyrosine kinases (RTKs) consisting of four members EGFR, ErbB2, ErbB3 and ErbB4 [8]. Upon binding, receptor dimerization and clustering occur leading to activation of downstream signaling pathways, including PI3K, ERK1/2, JAK/STAT, β-catenin, and calcium signaling [8-10].

Recombinant EGF protein is a key component of many organoid media, along with other stem cell niche factors, R-spondin 1, Noggin or Gremlin, Wnt3a and FGF10. In addition, recombinant EGF is used for establishing and maintaining intestine, stomach, liver, pancreas, brain and cancer organoids [11].

EGF has been implicated in several processes. These include angiogenesis where it has been shown to contribute to the proliferation of mesoderm-derived cells, epithelial development, cancer progression and inhibition of gastric secretion [12-14]. In wound healing applications, the role of EGF has been widely explored and shown to play a crucial role in promoting the wound healing processes including the differentiation of stem cells towards the tissue-specific cell types [3]. Its use has also been extended to various medical and cosmetic applications such as the production of collagen and accelerating skin regeneration [3,15-16].

Because of its critical role in stem cell differentiation, porcine EGF has recently gained wider attention for its use in transformative cultivated meat technology. Cultivated meat technology involves the use of animal cells to produce artificial meat also referred to as lab-grown meat [17]. Porcine EGF is a crucial growth factor that has been used in cell culture medium for the culture, proliferation, and differentiation of meat stem cells [18-20]. The use of porcine EGF will further accelerate the development and optimization of cell line-specific media. A study by Choi et al., showed that porcine EGF was crucial in maintaining and stabilizing pig muscle stem cells over a long period of time. The cultured cells maintained stemness evident from the expression of myogenic markers [18].

[1] Zhou S, Ochalek A, Szczesna K, et al. The positional identity of iPSC-derived neural progenitor cells along the anterior-posterior axis is controlled in a dosage-dependent manner by bFGF and EGF. Differentiation. 2016;92(4):183-194. doi: 10.1016/j.diff.2016.06.002

[2] Wolf CJ, Belair DG, Becker CM, Das KP, Schmid JE, Abbott BD. Development of an organotypic stem cell model for the study of human embryonic palatal fusion. Birth Defects Res. 2018;110(17):1322-1334. doi: 10.1002/bdr2.1394

[3] Basal O, Atay T, Ciris İM, Baykal YB. Epidermal growth factor (EGF) promotes bone healing in surgically induced osteonecrosis of the femoral head (ONFH). Bosn J Basic Med Sci. 2018;18(4):352-360. doi: 10.17305/bjbms.2018.3259

[4] Harris RC, Chung E, Coffey RJ. EGF receptor ligands. Exp Cell Res. 2003;284(1):2-13. doi: 10.1016/s0014-4827(02)00105-2

[5] Carpenter G, Cohen S. Epidermal growth factor. J Biol Chem. 1990;265(14):7709-7712. https://www.jbc.org/article/S0021-9258(19)38983-5/pdf

[6] Bell GI, Fong NM, Stempien MM, et al. Human epidermal growth factor precursor: cDNA sequence, expression in vitro and gene organization. Nucleic Acids Res. 1986;14(21):8427-8446. doi: 10.1093/nar/14.21.8427

[7] Carpenter G, Zendegui JG. Epidermal growth factor, its receptor, and related proteins. Exp Cell Res. 1986;164(1):1-10. doi: 10.1016/0014-4827(86)90449-0

[8] Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW. Epidermal growth factor receptor: mechanisms of activation and signalling. Exp Cell Res. 2003;284(1):31-53. doi: 10.1016/s0014-4827(02)00098-8

[9] Gamett DC, Pearson G, Cerione RA, Friedberg I. Secondary dimerization between members of the epidermal growth factor receptor family. J Biol Chem. 1997;272(18):12052-12056. doi: 10.1074/jbc.272.18.12052

[10] Qian X, O’Rourke DM, Fei Z, Zhang HT, Kao CC, Greene MI. Domain-specific interactions between the p185(neu) and epidermal growth factor receptor kinases determine differential signaling outcomes. J Biol Chem. 1999;274(2):574-583. doi: 10.1074/jbc.274.2.574

[11] Yin L, Li J, Zhang Y, et al. Changes in progenitors and differentiated epithelial cells of neonatal piglets. Anim Nutr. 2022;8(1):265-276. doi: 10.1016/j.aninu.2021.10.008

[12] Gospodarowicz D, Greenburg G, Bialecki H, Zetter BR. Factors involved in the modulation of cell proliferation in vivo and in vitro: the role of fibroblast and epidermal growth factors in the proliferative response of mammalian cells. In Vitro. 1978;14(1):85-118. doi: 10.1007/BF02618177

[13] Fisher DA. Epidermal growth factor in the developing mammal. Mead Johnson Symp Perinat Dev Med. 1988;(32):33-40. PMID: 2485437

[14] Hao C, Li Z, Zhang X, et al. Expression and clinical significance of EGF and TGF-α in chronic pancreatitis and pancreatic cancer. Minerva Endocrinol. 2018;43(3):253-258. doi: 10.23736/S0391-1977.17.02721-3

[15] Lee JH, Chellasamy G, Yun K, Nam MJ. EGF-expressed human mesenchymal stem cells inhibit collagenase1 expression in keratinocytes [published online ahead of print, 2023 Jul 26]. Cell Signal. 2023;110827. doi: 10.1016/j.cellsig.2023.110827

[16] Altuntas S, Dhaliwal HK, Eid Radwan A, Amiji M, Buyukserin F. Local epidermal growth factor delivery using nanopillared chitosan-gelatin films for melanogenesis and wound healing. Biomater Sci. 2022;11(1):181-194. Published 2022 Dec 20. doi: 10.1039/d2bm00836j

[17] Gomez Romero S, Boyle N. Systems biology and metabolic modeling for cultivated meat: A promising approach for cell culture media optimization and cost reduction. Compr Rev Food Sci Food Saf. 2023;22(4):3422-3443. doi: 10.1111/1541-4337.13193

[18] Choi KH, Yoon JW, Kim M, et al. Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro. Food Sci Anim Resour. 2020;40(4):659-667. doi: 10.1111/1541-4337.13193

[19] Lei Q, Ma J, Du G, Zhou J, Guan X. Efficient expression of a cytokine combination in Saccharomyces cerevisiae for cultured meat production. Food Res Int. 2023;170:113017. doi: 10.1016/j.foodres.2023.113017

[20] Choi KH, Kim M, Yoon JW, et al. Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29). Food Sci Anim Resour. 2020;40(5):852-859. doi: 10.5851/kosfa.2020.e51

FAQ

Porcine EGF (Epidermal Growth Factor) is a protein that belongs to the EGF family of proteins.

Porcine EGF is found in various tissues, including the gastrointestinal tissues.

Yes, porcine EGF is considered a cytokine that regulates various signalling pathways.

The porcine EGF gene encodes the protein responsible for promoting cell proliferation, differentiation, and survival.

Porcine EGF binds to the EGF receptor (EGFR) or ErbB1, a receptor tyrosine kinase.

The porcine EGF receptor, when activated by binding with EGF, triggers intracellular signalling pathways involved in cell growth, proliferation, and differentiation.

The porcine EGF pathway refers to the signalling cascade initiated upon binding of EGF to its receptor (EGFR), leading to activation of downstream signalling pathways such as the MAPK/ERK pathway and the PI3K/Akt pathway.

Porcine EGF is commonly used as a supplement in cell culture media to promote cell proliferation and survival, especially in cultures of stem cells, epithelial cells and organoids. It helps in maintaining cell viability and stimulating cell growth in vitro.

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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