Recombinant FGF2-G3 protein (Qk053)
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-free (AOF), carrier protein-free and with no His-tag.
Please note, FGF2-G3 will now be shipped in a lyophilized form. This change has been made to ensure we ship products in the most sustainable way possible, bringing FGF2-G3 in line with the rest of our portfolio. Please be assured that this change has no impact on bioactivity or quality. Please visit our reconstitution page for further information or contact us if you have any questions.
Recombinant FGF2-G3 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells. EC50 = 90 pg/ml (5.2 pM).
Cells are treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. Data from Qk053 lot #104340.
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.
We are a company founded and run by scientists to support innovation in stem cell biology and regenerative medicine. To enhance reliability and reproducibility in your applications, all our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.
FGF2-G3 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells.
Cells are treated in triplicate with a serial dilution of FGF2-G3 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity.
+ 72 hours
FGF2-G3 (Qk053) WT FGF2 (Qk027)
WT FGF-2 (Qk027) and FGF2-G3 (Qk053) were diluted in conditioned media and incubated at 37oC. Samples were taken at 24 h intervals and 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 (F/R).
Recombinant FGF2-G3 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 (Ludwig et al. 2006), StemPRO (Wang et al. 2007) and E8 (Chen et al. 2011). 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 interface. This thermostable FGF-2 is known as FGF2-G3, or FGF2-STAB® (Dvorak et al. 2017). 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 (Dvorak et al. 2017).
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 (Kuo et al. 2020 and updated in Lyra-Leite et al. 2020).
To manufacture and provide recombinant FGF2-G3 for stem cell culture, including use in B8 media and emerging applications such as cellular agriculture, 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 the academic forms of the protein for FGF2-G3 (Qk053), as these may cause issues for scientists looking to translate discoveries to clinical or scale-up applications, and introduce unnecessary scientific uncertainties.
Two forms of FGF-2 are used frequently in stem cell culture: 145 aa Qk025 and 154 aa (Qk027).
The 9 aa pro-segment of FGF2 present in the 154 aa FGF-2 is not required for biological activity and is thought to have roles in the localization of FGF-2 in vivo.
We wanted to allow scientists to compare directly with their existing FGF-2, so we introduced the nine amino acid substitutions into the 145 aa form of FGF-2 to make Qk052 FGF2-G3 (145 aa). The EC50 of the two forms of FGF2-G3 are 90 pg/ml and 47 pg/ml for the 154 aa and 145 aa forms respectively. As observed previously, the half-life is extended to >3 days.
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
What others are saying
There are no contributions yet.