Expression of recombinant Growth Factors using plant-based biorreactors: a comparative approach

MÜLLER, CAROLINA; MIRKIN, FEDERICO GABRIEL; PEREZ CASTRO, CAROLINA; WIRTH, SONIA ALEJANDRA; SEGRETIN, MARÍA EUGENIA

Congreso; XXXII Reunión Argentina de Fisiología Vegetal y XV Congreso Latinoamericano de Fisiología Vegetal; 2018

Growth factors are paracrine signaling proteins withmitotic and angiogenic activities which are also involved in the maintenance of the pluripotency state inhuman Embryonic Stem Cells(hESC). Given theimportanceof hESCsin regenerative medicine and research,the aim of this work is to develop different strategiesfor the production of human recombinant growth factors(rGFs) using plant-based biorreactors. Chloroplasttransformation allows the expression of high levels ofrecombinant proteinsin plants, and wasthereforechosen as our first approach to produce human FibroblastGrowth Factor (hFGFb). We cloned a 6xhis-tagged versionof hFGFb in a plastid transformation vector (pUTR-hFGFb)which was used totransformNicotiana tabacumleaves byparticle bombardment. PCR analysisofregenerated shootsconfirmed theintegration of thetransgen in fiveindependent transplantomiclines.Theselines are being subjectedtosuccessiveregeneration roundsto achieve homoplasmy.In addition, we are evaluating the production of hFGFb inNicotiana benthamiana plants by agroinfiltration, a transientexpression strategy. We adapted the pEAQ vector forrapid subcloning ofrecombinant proteinsfused to differentlocalization signals, to compare the accumulation of recombinant proteinsin differentsubcellularcompartments.Through thecomparison of thesestrategieseffectiveness,we attempt tooptimizerecombinant hFGFb