80 research outputs found
Design and validation of an immuno-PCR assay for IFN-α2b quantification in human plasma
Aim: Nowadays, IFN-α is considered a promising therapeutic target for systemic lupus erythematosus. An immuno-PCR (iPCR) was developed to quantify low amounts of IFN-α in human plasma followed by a deep analysis of the methodologic robustness throughout quality by design approach. Results: An accurate, sensitive, selective and versatile iPCR was validated. The critical iPCR procedural steps were identified, applying a Plackett-Burman design. Also, this assay demonstrated an outstanding LOD of 0.3 pg/ml. A significant aspect relies on its high versatility to detect and quantify other cytokines in human plasma as the appropriate biotinylated antibody is employed. Conclusion: This reliable iPCR assay can be clinically used as an alternative method for quantitating and detecting low IFN-α2b concentrations in human plasma samples.Fil: RodrĂguez, MarĂa Celeste. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Attallah, Carolina Veronica. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Lozano, Victoria. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Etcheverrigaray, Marina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Oggero Eberhardt, Marcos Rafael. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; Argentin
Novel erythropoietin-based therapeutic candidates with extra N-glycan sites that block hematopoiesis but preserve neuroplasticity
Neurological disorders affect millions of people causing behavior-cognitive disabilities. Nowadays they have no effective treatment. Human erythropoietin (hEPO) has been clinically used because of its neurotrophic and cytoprotective properties. However, the erythropoietic activity (EA) should be considered as a side effect. Some analogs like non-sialylated EPO, carbamylated EPO, or EPO peptides have been developed showing different weaknesses: erythropoiesis preservation, low stability, potential immunogenicity, or fast clearance. Herein, we used a novel strategy that blocks the EA but preserves hEPO neurobiological actions. N-glycoengineering was accomplished to add a new glycosylation site within the hEPO sequence responsible for its EA. hEPO-derivatives were produced by CHO.K1 cells, affinity-purified and functionally analyzed studying their in vitro and in vivo EA, their in vitro neuronal plasticity in hippocampal neurons and their neuroprotective action by rescuing hippocampal neurons from apoptosis. Muteins Mut 45_47 (K45 > N45 + N47 > T47), Mut 104 (S104 > N104), and Mut 151_153 (G151 > N151 + K153 > T153) lost their EA but preserved their neuroprotection activity and enhanced neuroplasticity more efficiently than hEPO. Interestingly, Mut 45_47 resulted in a promising candidate to explore as neurotherapeutic considering not only its biopotency but also its pharmacokinetic potential due to the hyperglycosylation.Fil: BĂŒrgi Fissolo, MarĂa de Los Milagros. Universidad Nacional del Litoral; Argentina. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; ArgentinaFil: Aparicio, Gabriela InĂ©s. Universidad Nacional de San MartĂn. Instituto de Investigaciones BiotecnolĂłgicas. - Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Oficina de CoordinaciĂłn Administrativa Parque Centenario. Instituto de Investigaciones BiotecnolĂłgicas; ArgentinaFil: Dorella, Aquiles. Universidad Nacional del Litoral; ArgentinaFil: Kratje, Ricardo Bertoldo. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral; ArgentinaFil: Scorticati, Camila. Universidad Nacional de San MartĂn. Instituto de Investigaciones BiotecnolĂłgicas. - Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Oficina de CoordinaciĂłn Administrativa Parque Centenario. Instituto de Investigaciones BiotecnolĂłgicas; ArgentinaFil: Oggero, Marcos. Universidad Nacional del Litoral; Argentin
Choice of the adequate quantification method for recombinant human GM-CSF produced in different host systems
Three enzyme-linked-immunosorbent assays (ELISA) were developed and
compared with a bioassay to quantify the recombinant human
granulocyte-macrophage colony stimulating factor (rhGM-CSF). These
assays were suitable to quantify the non-glycosylated rhGM-CSF present
in mixtures with variable protein content, and therefore useful for
determining concentrations of the cytokine in production processes.
Among these assays, the competitive ELISA, developed with a MAb that
recognises an epitope not involved in glycosylation, was the only one
suitable to quantify the glycosylated form of rhGM-CSF produced in
mammalian cell cultures
Neuroprotective activity of a new erythropoietin formulation with increased penetration in the central nervous system
Apart from its hematopoietic effect, erythropoietin (EPO) is a molecule with high neuroprotective potential. However, its prolonged application may cause serious adverse effects due to the erythropoiesis stimulation. Therefore, an EPO derivative with neuroprotective properties but low hematopoietic activity, designated as neuropoietin (rhNEPO), was developed in our lab using an alternative purification process of the recombinant human erythropoietic counterpart (rhEPO) produced in CHO cells [1]. The in vitro cytoprotective activity of rhNEPO on neural phenotype cells and its brain uptake from blood are herein analyzed.Fil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Ceaglio, Natalia Analia. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Mattio, MĂłnica Cristina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Roggero, Marcos Enrique. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Amadeo, Ignacio. Zelltek S. A.; ArgentinaFil: Forno, Angela Guillermina. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Perotti, Norma Maria. Zelltek S. A.; ArgentinaFil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de BioquĂmica y Ciencias BiolĂłgicas. Laboratorio de Cultivos Celulares; Argentin
Search for the lepton flavour violating decay tau(-) -> mu(-)mu(+)mu(-)
A search for the lepton flavour violating decay is performed with the LHCb experiment. The data sample corresponds to an integrated luminosity of 1.0 fb of proton-proton collisions at a centre-of-mass energy of 7 TeV and 2.0 fb at 8 TeV. No evidence is found for a signal, and a limit is set at 90% confidence level on the branching fraction, .A search for the lepton flavour violating decay Ï â ÎŒ ÎŒ ÎŒ is performed with the LHCb experiment. The data sample corresponds to an integrated luminosity of 1.0 fb of proton-proton collisions at a centre-of-mass energy of 7 TeV and 2.0 fb at 8 TeV. No evidence is found for a signal, and a limit is set at 90% confidence level on the branching fraction, .A search for the lepton flavour violating decay is performed with the LHCb experiment. The data sample corresponds to an integrated luminosity of of proton-proton collisions at a centre-of-mass energy of and at . No evidence is found for a signal, and a limit is set at confidence level on the branching fraction,
A study of CP violation in B-+/- -> DK +/- and B-+/- -> D pi(+/-) decays with D -> (KSK +/-)-K-0 pi(-/+) final states
A first study of CP violation in the decay modes and , where labels a or meson and labels a or meson, is performed. The analysis uses the LHCb data set collected in collisions, corresponding to an integrated luminosity of 3 fb. The analysis is sensitive to the CP-violating CKM phase through seven observables: one charge asymmetry in each of the four modes and three ratios of the charge-integrated yields. The results are consistent with measurements of using other decay modes
Study of forward Z + jet production in pp collisions at âs=7 TeV
A measurement of the +jet production cross-section in collisions at a centre-of-mass energy TeV is presented. The analysis is based on an integrated luminosity of recorded by the LHCb experiment. Results are shown with two jet transverse momentum thresholds, 10 and 20 GeV, for both the overall cross-section within the fiducial volume, and for six differential cross-section measurements. The fiducial volume requires that both the jet and the muons from the Z boson decay are produced in the forward direction (). The results show good agreement with theoretical predictions at the second-order expansion in the coupling of the strong interaction.A measurement of the +jet production cross-section in collisions at a centre-of-mass energy TeV is presented. The analysis is based on an integrated luminosity of recorded by the LHCb experiment. Results are shown with two jet transverse momentum thresholds, 10 and 20 GeV, for both the overall cross-section within the fiducial volume, and for six differential cross-section measurements. The fiducial volume requires that both the jet and the muons from the Z boson decay are produced in the forward direction (). The results show good agreement with theoretical predictions at the second-order expansion in the coupling of the strong interaction
Measurement of the (eta c)(1S) production cross-section in proton-proton collisions via the decay (eta c)(1S) -> p(p)over-bar
The production of the state in proton-proton collisions is probed via its decay to the final state with the LHCb detector, in the rapidity range GeV/c. The cross-section for prompt production of mesons relative to the prompt cross-section is measured, for the first time, to be at a centre-of-mass energy TeV using data corresponding to an integrated luminosity of 0.7 fb, and at TeV using 2.0 fb. The uncertainties quoted are, in order, statistical, systematic, and that on the ratio of branching fractions of the and decays to the final state. In addition, the inclusive branching fraction of -hadron decays into mesons is measured, for the first time, to be , where the third uncertainty includes also the uncertainty on the inclusive branching fraction from -hadron decays. The difference between the and meson masses is determined to be MeV/c.The production of the state in proton-proton collisions is probed via its decay to the final state with the LHCb detector, in the rapidity range . The cross-section for prompt production of mesons relative to the prompt cross-section is measured, for the first time, to be at a centre-of-mass energy using data corresponding to an integrated luminosity of 0.7Â fb , and at using 2.0Â fb . The uncertainties quoted are, in order, statistical, systematic, and that on the ratio of branching fractions of the and decays to the final state. In addition, the inclusive branching fraction of -hadron decays into mesons is measured, for the first time, to be , where the third uncertainty includes also the uncertainty on the inclusive branching fraction from -hadron decays. The difference between the and meson masses is determined to be .The production of the state in proton-proton collisions is probed via its decay to the final state with the LHCb detector, in the rapidity range GeV/c. The cross-section for prompt production of mesons relative to the prompt cross-section is measured, for the first time, to be at a centre-of-mass energy TeV using data corresponding to an integrated luminosity of 0.7 fb, and at TeV using 2.0 fb. The uncertainties quoted are, in order, statistical, systematic, and that on the ratio of branching fractions of the and decays to the final state. In addition, the inclusive branching fraction of -hadron decays into mesons is measured, for the first time, to be , where the third uncertainty includes also the uncertainty on the inclusive branching fraction from -hadron decays. The difference between the and meson masses is determined to be MeV/c
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