PromoKine's animal-free cytokines and growth factors have been produced by molecular farming using barley grain or tobacco plants as production host - which has also the G.R.A.S. (Generally Recognized As Safe) status from the FDA.
These cytokines are free of animal components, serum, endotoxins and antibiotics as well as human or animal infectious agents or other endogenous mammalian contamination (e.g. traces of unwanted growth factors and cytokines). As these proteins have been expressed in barley seed endosperm or tobacco plants, they show very low proteolytic, pyrogenic and inflammatory activity compared to other commercially available proteins produced in bacteria, yeast, insect or mammalian cells. In addition, they retain proper eukaryotic folding and glycosylation resulting in an increased stability and biological activity. Degradation due to protease carryover as well as unwanted cell responses (e.g. proliferation, chemokine release and inflammatory cascades) or cell growth inhibition caused by contaminating serum components (serum carryover), endotoxins or bacterial, viral and prion infectious agents are eliminated or at least extremely reduced.
Note: Endotoxins (lipopolysaccharides, LPS) are produced and released by Gram-negative bacteria. They are frequently contaminating protein preparations derived e.g. from E. coli and are difficult to remove. Cell culture experiments have shown that even low levels of endotoxins currently regarded as acceptable (~1 EU/µg) and found in many commercially available cytokines and growth factors can have profound effects on cell viability as well as cellular behaviour and functions. In particular, even low endotoxin traces found in many cytokines and growth factors produced in E. coli (indicating endotoxin contents <1 EU/µg) may be quite critical for stem cell experiments as final endotoxin concentrations of 0.001 ng/ml in the medium can already have significant effects on stem cell cultivation (e.g. failure to differentiate into functional cell types, general adverse effects on cell viability and proliferation). Endotoxin traces in E. coli-derived cytokines and growth factors are also biasing angiogenesis studies as they show considerable angiogenic effects inducing tumour cell proliferation and VEGF production and decreasing apoptosis.
PromoKine's animal-free cytokines and growth factors typically have an endotoxin level of less than 0.05 EU/µg (i.e. they are virtually free of endotoxins) excluding such complications in cell culture.
Note: PromoKine now also provide endotoxin-free HSA that can be used as carrier protein for long-term storage of cytokines and growth factors in solution.
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Figure 1: PromoKine's animal-free and endotoxin-free FGF-2 showed equivalent biological activity when compared to the respective, high-priced growth factor of a market leader.
| Figure 2: In a cell-based activity assay a PromoKine animal-free cytokine showed equivalent biological activity but significantly less cytotoxic effects than two GMP compliant cytokines from competitors. | Figure 3: Endotoxin Levels in Cytokines and Growth Factors. |
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Proteins / Signaling Molecules