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Recombinant FGF2-G3 (154aa) protein

QK053

Brand: Qkine

Recombinant FGF2-G3 (FGF2-STAB®) protein is a thermostable engineered form of FGF-2 (bFGF). Qk053 is the 154 aa mature domain of FGF-2 (Qk027) with nine amino acid substitutions to enhance stability without impacting bioactivity developed by Dvorak et al. 2018. This increases the functional half-life of the protein from <10 h (wild-type) to >7 days (FGF2-G3).

Recombinant FGF2-G3 is used in B8 media (Kuo et al. 2019) for weekend free, high homogeneity induced pluripotent stem cell culture. FGF2-G3 also has applications in chemically defined stem cell and organoid culture media, and cultured meat media development.

High purity 17 kDa bioactive FGF2-G3 protein, animal origin-free (AOF), carrier protein-free and tag free.

Currency: 

Product name Catalog number Pack size Price Price (USD) Price (GBP) Price (EUR)
Recombinant FGF2-G3 protein, 50 µg QK053-0050 50 µg (select above) $ 185.00 £ 140.00 € 164.00
Recombinant FGF2-G3 protein, 100 µg QK053-0100 100 µg (select above) $ 280.00 £ 210.00 € 246.00
Recombinant FGF2-G3 protein, 500 µg QK053-0500 500 µg (select above) $ 725.00 £ 540.00 € 631.00
Recombinant FGF2-G3 protein, 1000 µg QK053-1000 1000 µg (select above) $ 1,050.00 £ 800.00 € 935.00

Note: prices shown do not include shipping and handling charges.

Qkine company name and logo are the property of Qkine Ltd. UK.

Alternative protein names
Basic fibroblast growth factor, bFGF, FGF-β, FGF2, Fibroblast growth factor-basic, HBGF-2, FGF2-G3, FGF2-STAB, FGF2-STAB2
Species reactivity
Species neutral
Alternative(s)

Summary

Handling and Storage FAQ

Featured applications

  • Expansion of induced pluripotent, embryonic and mesenchymal stem cells

Bioactivity

Recombinant FGF2-G3 activity was determined using the Promega serum response element luciferase reporter assay in transfected HEK293T cells. EC50 = 90 pg/ml (5.2 pM). Cells were treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity was measured and normalized to the control Renilla luciferase activity. Data from Qk053 lot #104340.
 

Qk053-FGF2-G3-bioactivity-1

 

Purity

Recombinant FGF2-G3 migrates as a major band at 17 kDa in non-reducing (NR) conditions. The higher molecular weight minor band is the dimeric form. Upon reduction (R), only the 17 kDa band is visible. No contaminating protein bands are present. Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk053 batch #104340.

210920-SDS-PAGE-FGF2-G3-104340-3-ug

 

Further quality assays

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

Qk027-vs-FGF2-G3-stability-1024x400

WT FGF-2 (Qk027) and FGF2-G3 (Qk053) were diluted in conditioned media and incubated at 37oC for 2 or 7 days before FGF-2 activity was assayed in triplicate using the Promega serum response element luciferase reporter assay in transfected HEK293T cells. Firefly luciferase activity was normalized to the control Renilla luciferase activity.


Protein background

FGF-2 (also known as basic FGF or bFGF) is an essential growth factor for maintaining human embryonic stem cell (hESC) and induced pluripotency stem cell (iPSC) pluripotency in feeder-free and chemically defined stem cell media. It is a core component of widely adopted media including mTESR [1, 2], StemPRO [3] and E8 [4]. However, FGF-2 is inherently unstable and prone to proteolytic degradation and aggregation. This fundamental biochemical instability, and therefore low half-life in culture media (<10 h), is an important contribution to the need for frequent media changes and challenges in improving homogeneity during stem cell proliferation and subsequent differentiation.

To improve the stability of FGF-2 for stem cell media and regenerative medicine applications, Dvorak and colleagues at Masaryk University used computer-assisted protein engineering to identify an optimal set of nine amino acid substitutions that stabilize FGF-2. These substitutions were designed to avoid structural changes to the FGF receptor 1 (FGFR1) and FGF receptor 2 (FGFR2) binding sites. This thermostable FGF-2 is known as FGF2-G3, or FGF2-STAB® [5]. The biological activity of wild-type FGF-2 is <50% after 10h incubation with conditioned media. In contrast, no reduction in FGF2-G3 biological activity is observed after >7 days incubation with conditioned media at 37oC. Both FGF2-G3 and wild-type FGF-2 maintain hESC pluripotency and expression of pluripotency markers Oct-4 and nanog with equivalent efficacy [5].

In 2020, Paul Burridge and colleagues at Northwestern University, Chicago, published a protocol for B8 media. This iPSC maintenance media uses thermostable FGF2-G3, along with optimization of media component concentration and composition to reduce media cost and facilitate weekend-free stem cell culture regimes [6, 7].

To manufacture and provide FGF2-G3 for stem cell culture, including use in B8 media and emerging applications such as cultured meat, Qkine has licensed the patented FGF2-G3 technology from Enantis/Masaryk University. We have combined the excellent science behind the FGF2-G3 technology with our protein manufacture expertise to provide gold standard protein for use in cell culture media. We have removed His tags present in academic forms of the protein, as these may give rise to issues for scientists translating discoveries to the clinical or scale-up. His tags also introduce unnecessary scientific uncertainty.

Background references

  1. Ludwig, T. E. et al. Derivation of human embryonic stem cells in defined conditions. Nat. Biotechnol. 24, 185–187 (2006). doi.org/10.1038/nbt1177

  2. Ludwig, T. E. et al. Feeder-independent culture of human embryonic stem cells. Nat. Methods 3, 637–646 (2006). doi.org/10.1038/nmeth902

  3. Wang L, Schulz TC, Sherrer ES et al. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling. Blood. 2007 Dec 1;110(12):4111-9. doi: 10.1182/blood-2007-03-082586.

  4. Chen G, Gulbranson DR, Hou Z et al. Chemically defined conditions for human iPSC derivation and culture. Nat Methods. 2011 May;8(5):424-9. doi: 10.1038/nmeth.1593.

  5. Dvorak P, Bednar D, Vanacek P, et al. Computer-assisted engineering of hyperstable fibroblast growth factor 2. Biotechnology and Bioengineering. 2018; 115: 850–862. https://doi.org/10.1002/bit.26531

  6. Kuo H H, Gao X, DeKeyser J-M, K. Fetterman A, et al. Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture, Stem Cell Reports 2020 https://doi.org/10.1016/j.stemcr.2019.12.007

  7. Lyra-Leite D M, Fonoudi H, Gharib M, Burridge P W. An updated protocol for the cost-effective and weekend-free culture of human induced pluripotent stem cells, STAR Protocols 2021. https://doi.org/10.1016/j.xpro.2020.100213


Publications using Recombinant FGF2-G3 (154 aa) protein (Qk053)


FAQ

What is FGF-2?
Fibroblast growth factor 2 (FGF-2), also known as basic fibroblast growth factor (bFGF) is a growth factor and signaling protein.

Where is FGF-2 found?
FGF-2 is expressed in a developmental and tissue specific manner. It’s expression is tightly controlled in normal tissues and it can be detected in all major tissues.

What does FGF-2 do?
FGF-2 is essential for normal embryonic development. It has roles in cell survival and proliferation, angiogenesis, tumorigenesis, wound healing and tissue repair.

What does FGF-2 bind to?
FGF-2 binds to and signals though all four of the FGF receptors FGFR1-4.

What is the function of the FGF receptor?
FGFRs phosphorylate specific tyrosine residues and activate the RAS-MAPK, PI3K-AKT, PLCγ, and STAT intracellular signaling pathways.

How is FGF-2 used in stem cell culture?
FGF-2 is used to maintain the pluripotency of stem cells in culture.