What is Biorisk-free growth factor - Isokine™? Animal-free Serum-free ORF Genetics has developed a revolutionary production system for biorisk-free recombinant proteins of animal origin. The Orfeus™ system uses barley endosperm storage cell, characterised by its biochemically inert environment and lack of endotoxins, as the production host. Further advantages of this system include low protease activity and secondary metabolite content and simple protein content all of which aid in downstream processing. Barley has also the G.R.A.S. (Generally Recognised As Safe) status from FDA. The Orfeus™ system is used to produce ORF’s Isokine™ line of growth factors and cytokines for biomedical research. Endotoxin-free Endotoxin levels (turbidimetric kinetic assay) 1.2 1 EU/ug 1 0.8 0.6 0.4 0.2 0.00135 0 0.002 VEGF121 ISOkine IFN-gamma ISOkine IFN-gamma commercial* Ref. Associates of Cape Cod Industries, Deacon Park, Knowsley, Liverpool, UK *E.coli production; company´s claim: "Less than 1 EU/ug of recombinant IFN gamma" Low protease activity The barley endosperm tissue has a very low protease activity in comparison to most other cellular organisms, resulting in the unique protein storage function of the seed, that can maintain the proteins, including recombinant proteins, intact for years. Protease activity (azocasein protein activity assay)** Protease activity (azocasein protein activity assay)* 9.45 10 dA440/hr*mg 8 6 4 2 0 0.56 0.6 0.4 0.3 0.2 0.1 0 -0.1 0.2 barley extract -0.2 E.coli BL21 -0.07 endospern extract human cells Ref. Dept. of Biochemistry, Science Institute, University of Iceland, Reykjavik, Iceland *100mM Tris pH8.0; **100mM MOPS pH 6. Orfeus™ The next level of assurance Low pyrogenic and pro-inflammatory activity Induction of IL-6 in Mononuclear cells* (monocyte activation test; MAT) 12000 10000 8000 pg/ml 6000 4000 2000 0 Barley background GF blank Neg. control Pos. control Ref. The Blood Bank, University hospital, Reykjavik, Iceland *TNA-alpha and IL-1beta also tested with similar results Lectin analysis on Western blot* 1 2 3 4 1 endosperm barley extract 2 flowthrough from column A 3 IGF-1 elute from column A 4 IGF-1 elute from column B *wheat germ agglutinin antibody downstream processing and purification easier and less technically demanding. The water and salt soluble fraction of barley endosperm proteins accounts for 21% of the total proteins. For water soluble recombinant proteins, such as the ISOkines™, already 79-80% of the seed proteins can separated from the protein of interest by simple means of extraction. Orfeus™ The next level of assurance ORF Genetics Keldnaholt IS-112 Reykjavik Iceland phone +354 591 1570 fax +354 591 1580 [email protected] www.orfgenetics.com N-America phone 1 800 705 7408 fax 1 800 494 3294 80749 The relatively simple protein profile of the endosperm storage tissue makes • Simple protein profile SÍA as acknowledged by regulatory authorities. • Plant tissue is not a source of any known human- or animal infectious agents, PI PAR Void of human or animal infectious agents