title: Transgenic tobacco plants as production platform for biologically active human interleukin 2 and its fusion with proteinase inhibitors.
abstract: Transgenic plants offer a low-cost approach for the production of pharmaceutically important and commercially valuable recombinant proteins. Our studies were focused on the plant-based production of human interleukin 2 (hIL-2) and its fusion with proteinase inhibitors, either SPI2 from Galleria mellonella or CMTI from Cucurbita maxima. Finally, five plant expression cassettes were obtained. Three of them contained the single cDNA encoding CMTI I, SPI2 and hIL-2, respectively, while two of them contained the translational fusion, SPI2::hIL-2 and CMTI::hIL-2. In all cases, the transgenes were controlled by the RbcS1 promoter and terminator and the recombinant proteins were targeted to the endoplasmic reticulum. After tobacco transformation, five groups of transgenic plants were obtained and analysed. The level of recombinant proteins was estimated either by Western blot or by ELISA. The biological activity of plant-produced hIL-2 alone or in a fusion with SPI2 or CMTI was confirmed using the mammalian cells proliferation assay. The activities of proteinase inhibitors were confirmed in proteolysis assay using azocoll as a substrate. The usefulness of using proteinase inhibitor CMTI I in a fusion with hIL-2 as a protective agent against trypsin digestion was demonstrated.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: Patrycja
authors.initials: P
authors.lastname: Redkiewicz
authors.forename: Aneta
authors.initials: A
authors.lastname: Więsyk
authors.forename: Anna
authors.initials: A
authors.lastname: Góra-Sochacka
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Sirko
mesh_terms: D001709:Biotechnology / Q000379:methods*; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis*; D030821:Plants, Genetically Modified; D011480:Protease Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic; D014357:Trypsin / Q000378:metabolism
chemical_list: C508594:IL2 protein, human; D007376:Interleukin-2; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins; D014357:Trypsin