Protein design and immunogenic analysis of COVID-19 vaccine candidates based on RBD/trimeric-spike antigens and chimeric VLPs antigens

Spike glycoprotein (S) and its Receptor Binding Domain region (RBD) are the main targets of neutralizing antibodies during an infection with SARS-CoV-2, the etiologic agent of COVID-19 pandemic viral disease. Thus, they are the chosen antigens for the development of diagnostic kits and vaccines candidates. Furthermore, Virus-like Particles (VLPs) constitute potent immunogens that have been engineered to obtain vaccine candidates through expression of SARS-CoV-2 S, M, E and N proteins. Although it might be a challenging platform, as multiple proteins must be expressed in order to assure VLP budding, they are able to induce stronger immune responses than soluble antigens due to their particulate nature and highly repetitive antigen display. Hence, we aimed to generate a serum-free platform to produce soluble SARS-CoV-2 antigens and design a novel chimeric VLP (cVLP) exposing the prefusion stabilized S ectodomain on its surface. Please click Download on the upper right corner to see the full abstract

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 Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Ceaglio, Natalia Analia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Mattio, Mónica Cristina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Roggero, Marcos Enrique. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Amadeo, Ignacio. Zelltek S. A.; ArgentinaFil: Forno, Angela Guillermina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Perotti, Norma Maria. Zelltek S. A.; ArgentinaFil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentin

Aprendizajes y prácticas educativas en las actuales condiciones de época: COVID-19

“Esta obra colectiva es el resultado de una convocatoria a docentes, investigadores y profesionales del campo pedagógico a visibilizar procesos investigativos y prácticas educativas situadas en el marco de COVI-19. La misma se inscribe en el trabajo llevado a cabo por el equipo de Investigación responsable del Proyecto “Sentidos y significados acerca de aprender en las actuales condiciones de época: un estudio con docentes y estudiantes de la educación secundarias en la ciudad de Córdoba” de la Facultad de Filosofía y Humanidades. Universidad Nacional de Córdoba. El momento excepcional que estamos atravesando, pero que también nos atraviesa, ha modificado la percepción temporal a punto tal que habitamos un tiempo acelerado y angustiante que nos exige la producción de conocimiento provisorio. La presente publicación surge como un espacio para detenernos a documentar lo que nos acontece y, a su vez, como oportunidad para atesorar y resguardar las experiencias educativas que hemos construido, inventado y reinventando en este contexto. En ella encontrarán pluralidad de voces acerca de enseñar y aprender durante la pandemia. Este texto es una pausa para reflexionar sobre el hacer y las prácticas educativas por venir”.Fil: Beltramino, Lucia (comp.). Universidad Nacional de Córdoba. Facultad de Filosofía y Humanidades. Escuela de Archivología; Argentina

Structure and properties of biocompatible poly(glycerol adipate) elastomers modified with ethylene glycol

Polyesters poly(glycerol adipate)s or PGAs were prepared through a catalyst-free polycondensation reaction involving different proportions of adipic acid and glycerol. Additionally, another set of PGAs was formulated by adding ethylene glycol to the reaction mixture, creating three PGA-co-ethylene glycols of different ethylene glycol-to-glycerol ratios. All prepolymers were chemically crosslinked into elastomeric films according to specific experimental conditions. A full characterization of elastomers revealed an ease of tunability according to monomer ratios and curing conditions. Mechanical properties presented a variety of Young's modulus values ranging from 0.07 to 8.33 MPa. Within this range, a number of Young's modulus values of soft tissues can be included. Moreover, a cytotoxicity analysis performed on embryonic mouse fibroblasts (NIH/3T3) showed an absence of cytotoxicity and good adherence over all PGA and PGA-co-ethylene glycol elastomers. In addition, fibroblasts underwent a differentiation process into neuron-like cells, thus exhibiting excellent potential for soft tissue engineering.Fil: Navarro, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Ceaglio, Natalia Analia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Rintoul, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentin

Effect of ANITVNITV peptide fusion on the bioactivity and pharmacokinetics of human IFN-α2b and a hyper-N-glycosylated variant

Different strategies have been developed and successfully applied to biotherapeutics in order to improve their in vivo efficacy. The genetic fusion to natural or synthetic glycosylated peptides constitutes a promising strategy since it conserves the protein sequence and results in the improvement of the pharmacokinetic properties. The ANITVNITV peptide described by Perlmann and coworkers presents 9 amino acids and 2 potential N-glycosylation sites. Its fusion to FSH resulted in the increase of the molecular mass and negative charge of the protein. Consequently, the pharmacokinetics was considerably improved. The aim of the present study was to compare the influence of ANITVNITV peptide fusion on the physicochemical, biological and pharmacokinetic properties of native hIFN-α2b (IFNwt), which contains a single O-glycosylation site, and a hyperglycosylated variant (IFN4N), that bears, in addition, 4 N-linked glycans. The resulting molecules, IFNwtNter and IFN4NNter, evidenced a higher molecular mass and negative charge compared to IFNwt and IFN4N, respectively. Therefore, the pharmacokinetic properties of the new molecules were significantly improved. The molecules obtained by the synthetic peptide fusion strategy evidenced a decrease in their in vitro antiviral specific biological activities (SBA). However, in vitro antiproliferative SBA was differentially modified for IFNwtNter and IFN4NNter in comparison with the parental molecules. For IFNwtNter, a reduction in the antiproliferative SBA was also observed. Remarkably, the addition of the ANITVNITV peptide to the N-terminus of IFN4N had a positive impact on its growth-inhibitory activity. This feature together with its improved pharmacokinetics encourages the development of IFN4NNter as an IFN-α based biobetter.Fil: Gugliotta, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Ceaglio, Natalia Analia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Kratje, Ricardo Bertoldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Roggero, Marcos Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentin

Influence of carbohydrates on the stability and structure of a hyperglycosylated human interferon alpha mutein

Protein physical and chemical instability is one of the major challenges in the development of biopharmaceuticals during every step of the process, ranging from production to final delivery. This is particularly applicable to human recombinant interferon alpha-2b (rhIFN-α2b), a pleiotropic cytokine currently used worldwide for the treatment of various cancer and chronic viral diseases, which presents a poor stability in solution. In previous studies, we have demonstrated that the introduction of four N-glycosylation sites in order to construct a heavily glycosylated IFN variant (4N-IFN) resulted in a markedly prolonged plasma half-life which was reflected in an enhanced therapeutic activity in mice in comparison with the commercial non-glycosylated rhIFN-α2b (NG-IFN). Herein, we evaluated the influence of glycosylation on the in vitro stability of 4N-IFN towards different environmental conditions. Interestingly, the hyperglycosylated cytokine showed enhanced stability against thermal stress, acid pH and repetitive freeze-thawing cycles in comparison with NG-IFN. Besides, microcalorimetric analysis indicated a much higher melting temperature of 4N-IFN, also demonstrating a higher solubility of this variant as denoted by the absence of precipitation at the end of the experiment, in contrast with the NG-IFN behaviour. Furthermore, far-UV circular dichroism (CD) spectrum of 4N-IFN was virtually superimposed with that of NG-IFN, indicating that the IFN structure was not altered by the addition of carbohydrate moieties. The same conclusion could be inferred from limited proteolysis studies. Our results suggest that glycoengineering could be a useful strategy for protecting rhIFN-α2b from inactivation by various external factors and for overcoming aggregation problems during the production process and storage.Fil: Ceaglio, Natalia Analia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Oggero Eberhardt, Marcos Rafael. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Poly-(pentaerythritol adipate)-co-(poly ethylene glycol) Elastomers for the Sustained Release of Paclitaxel

Over the past 10 years poly-(polyol alkanoate)s elastomers have intensively been investigated due to their extraordinary potential for soft tissue engineering applications. In this work, a family of novel hyperbranched elastomers based on pentaerythritol and adipic acid, modified with PEG of 200 and 400 Da, is synthesized. The polymers are obtained by a simple 2 steps thermal process that avoids the use of toxic catalysts. Noteworthy, elastomers properties can be finely tuned by adjusting the molar ratio of monomers and curing conditions. The elastomers, which are fully characterized by standard methods, show excellent results in the in vitro controlled release of Paclitaxel. Cell culture assays indicate that all materials are able to inhibit lung cancer cell proliferation and that the release of Paclitaxel reduce cancer cell viability, indicating that the elastomers prepared hereby are good candidates for their use as eluting systems in long term cancer treatment.Fil: Navarro, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Minari, Roque Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Ceaglio, Natalia Analia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Masin, Marianela. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Vaillard, Santiago Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentin

Novel poly(diol sebacate)s as additives to modify paclitaxel release from poly(lactic-co-glycolic acid) thin films

Paclitaxel (PTX) incorporation in poly (lactic acid-co-glycolic acid) (PLGA) matrices produce films with high tensile rigidity and slow release that fail to deliver the required release rate for most biomedical applications such as in drug eluting stents and cancer treatments. To modify and improve this behavior, a set of poly (diol sebacate)s using different aliphatic diol length (1,3propanediol, 1,9-nonanediol and 1,10-decanediol) were synthesized and fully characterized as possible additives. The tensile properties of PLGA blends were evaluated as these materials could be used as drug eluting stent coatings. A significant improvement of mechanical flexibility was observed with 20 % additive content, as it reduced the Young´s Modulus (YM) value and increased the maximum deformation at break. PTX release was studied and correlated with the release of additive from PLGA films. An increase of the initial burst release phase was observed on all the blends when compared to the control films of PLGA. Modulation of PTX release was achieved by altering the hydrophilicity degree of the additive or its percentage content on the blend. This supports the possibility that PTX was partitioned into the additive phase. Cytotoxicity analyses of novel additives were performed on mouse embryonic fibroblasts NIH/3T3.Fil: Navarro, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Mogosanu, Diana Elena. University of Ghent; BélgicaFil: Ceaglio, Natalia Analia. Universidad Nacional del Litoral; ArgentinaFil: Luna, Julio Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; ArgentinaFil: Dubruel, Peter. University of Ghent; BélgicaFil: Rintoul, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química. Universidad Nacional del Litoral. Instituto de Desarrollo Tecnológico para la Industria Química; Argentin