Recombinant human/ bovine/ porcine TGF-β1 PLUS™ protein

Summary

• High purity optimised TGF-β1 protein (Uniprot: P01137)
• 24 kDa (dimer)
• >98%, by SDS-PAGE quantitative densitometry
• Expressed in E. coli
• Animal origin-free (AOF) and carrier protein-free
• Manufactured in Qkine's Cambridge, UK laboratories
• Lyophilized from acetonitrile, TFA
• Resuspend in 10 mM HCl (Reconstitution solution A) at >50 µg/ml, add carrier protein if desired, prepare single-use aliquots and store frozen at -20 °C (short-term) or -80 °C (long-term)

Featured applications

• iPSC and ESC maintenance and expansion
• Chemically defined media

Purity

Purity

Protein background

Transforming growth factor-beta 1 (TGF-β1) is a pleiotropic cytokine part of the TGF-β superfamily. TGF-β1 regulates various cellular processes, including cell proliferation, growth, differentiation, motility, and apoptosis [1]. It plays a crucial role in the immune response, tissue repair, and the epithelial-mesenchymal transition. Transforming growth factor-beta 1 is produced by various cell types, including immune cells, fibroblasts, and epithelial cells. It is synthesized and secreted as an inactive or latent complex, associated with latency-associated proteins (LAP), and targeted to the extracellular matrix [2]. It is released from latency by TGF-β activators including plasmin, matrix metalloproteases, integrins. Once the LAP cleaved, the mature transforming growth factor-beta 1 is a homodimeric protein composed of two identical subunits linked by a disulfide bond. Its amino acid sequence is composed of 390 amino acids. TGF-β1 signals through complexes of cell surface receptors including TGF-βRII/TGF-βRI and ALK-5/ALK-1. This triggers downstream signaling cascades, such as the Smad-dependent and Smad-independent pathways.

In cell culture, recombinant TGF-β1 protein is an essential growth factor in many embryonic and induced pluripotent stem cell maintenance media, including the commonly used chemically-defined E8, StemPro, and mTeSR medias [3–5]. Transforming growth factor-beta 1 supports the survival and maintenance of pluripotency of stem cells [1]. TGF-β1 is used to promote the differentiation of various cell types such as fibroblasts, epithelial cells, and immune cells. It is used in combination with other growth factors such as BMP-2 to regulate bone marrow stromal cell differentiation or with IL-2 and IL-6 to regulate T reg and Th17 cells differentiation [6–8].

To date, recombinant human TGF-β1 has only been produced from mammalian cell protein expression systems (HEK or CHO), where endogenous protein contaminants, cost and animal-free status is a challenge. As part of the ongoing mission to redefine industry standards for growth factor and cytokine biochemical quality, Qkine introduced the first optimized, animal-free, and highly bioactive recombinant human TGF-β1.

Benefits:

• Truly animal-free alternative for chemically-defined stem cell culture media
• High potency in iPSC culture with high pluripotency (nanog) marker expression
• High purity, extensive biochemical data, and exceptional lot-to-lot consistency

Background references

1. D. James, A. J. Levine, D. Besser, and A. Hemmati-Brivanlou, ‘TGFβ/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells’, Development, vol. 132, no. 6, pp. 1273–1282, Mar. 2005, doi: 10.1242/dev.01706.
2. J. P. Annes, J. S. Munger, and D. B. Rifkin, ‘Making sense of latent TGFβ activation’, J. Cell Sci., vol. 116, no. 2, pp. 217–224, Jan. 2003, doi: 10.1242/jcs.00229.
3. J. Beers et al., ‘Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions’, Nat. Protoc., vol. 7, no. 11, pp. 2029–2040, 2012, doi: 10.1038/nprot.2012.130.
4. T. E. Ludwig, V. Bergendahl, M. E. Levenstein, J. Yu, M. D. Probasco, and J. A. Thomson, ‘Feeder-independent culture of human embryonic stem cells’, Nat. Methods, vol. 3, no. 8, Art. no. 8, Aug. 2006, doi: 10.1038/nmeth902.
5. A. Wang et al., ‘Induced Pluripotent Stem Cells for Neural Tissue Engineering’, Biomaterials, vol. 32, no. 22, pp. 5023–5032, Aug. 2011, doi: 10.1016/j.biomaterials.2011.03.070.
6. Y. Tang et al., ‘TGF-β1–induced migration of bone mesenchymal stem cells couples bone resorption with formation’, Nat. Med., vol. 15, no. 7, Art. no. 7, Jul. 2009, doi: 10.1038/nm.1979.
7. M. Elsafadi et al., ‘Convergence of TGFβ and BMP signaling in regulating human bone marrow stromal cell differentiation’, Sci. Rep., vol. 9, no. 1, Art. no. 1, Mar. 2019, doi: 10.1038/s41598-019-41543-0.
8. M. Veldhoen, R. J. Hocking, C. J. Atkins, R. M. Locksley, and B. Stockinger, ‘TGFβ in the Context of an Inflammatory Cytokine Milieu Supports De Novo Differentiation of IL-17-Producing T Cells’, Immunity, vol. 24, no. 2, pp. 179–189, Feb. 2006, doi: 10.1016/j.immuni.2006.01.001.

Publications using Recombinant human TGF-β1 PLUS™ protein (Qk010)

Modeling the selective growth advantage of genetically variant human pluripotent stem cells to identify opportunities for manufacturing process control
Beltran-Rendon C, Price CJ, Glen K et al.
DOI:  10.1016/j.jcyt.2024.01.010

Refined home-brew media for cost-effective, weekend-free hiPSC culture and genetic engineering
Truszkowski L, Bottini S, Bianchi S et al.
DOI: https://doi.org/10.12688/openreseurope.18245.1

Recombinant production of growth factors for application in cell culture
Venkatesan M, Semper C, Skrivergaard S et al.
DOI: 10.1016/j.isci.2022.105054.

FAQ

What does TGF-beta 1 do?
TGF-β1 is a multifunctional cytokine that plays a crucial role in various cellular processes. It regulates cell growth, differentiation, proliferation, migration, and apoptosis. It is also involved in the immune response, epithelial-mesenchymal transition, and the extracellular matric synthesis and modelling.

What kind of protein is TGF-beta 1?
It is a member of the transforming growth factor-beta superfamily of cytokines that are synthesized as latent complexes. TGF-β1 requires further processing to become the biologically active, mature TGF-β1.

What diseases are associated with TGF-beta 1?
Dysregulation of TGF-β1 signaling is associated with various diseases such as cancer, fibrosis, autoimmune, cardiovascular, neurological, and chronic kidney diseases.

What does a high human TGF-beta 1 mean?
Elevated levels of TGF-β1 in the human body can have different implications such as higher risks of fibrosis, cancer progression, chronic inflammation, and cardiovascular diseases. It is important to consider the specific clinical context, the presence of other biomarkers, and the patient’s prognosis.

What activates TGF-beta 1?
TGF-β1 is synthesized and secreted in an inactive or latent form. It is activated through cleavage and binding to matrix metalloproteinases, integrins, extracellular matrix such as thrombospondin-1, but also pH changes and reactive oxygen species.

How is TGF-beta 1 used in cell culture?
It is an essential growth factor in many embryonic and induced pluripotent stem cell maintenance media, including the commonly used E8, StemPro, and mTeSR media. TGF-β1 also promotes the differentiation of various cell types such as fibroblasts, epithelial cells, and immune cells.

What is Qkine TGF-β1 ‘PLUS’?
Qkine TGF-β1 PLUS™ protein is the first entirely animal origin-free recombinant TGF-β1 protein, TGF-β family members are difficult to express in E. coli due to their complex folding so are only commercially available expressed in animal cells. Qkine have optimized the production of TGF-β1 to produce a pure animal origin-free protein or highly reproducible results and compatible with chemically-defined stem cell media.