Improving the downstream processing of interferon alfa-2b using alternative purification platforms based on ionic liquids

Improvements on human life expectancy and the lack of effective therapies has led to an increment of chronic diseases, being the application of biopharmaceuticals an efficient strategy to mitigate this scenario. Among the current available biopharmaceuticals, the role of interferon α-2b (IFNα-2b) should be highlighted, as it has been marketed over 30 years with a considerable impact on the global therapeutic proteins market (Castro et al, Vaccines, 2021). IFN manufacturing requires the use of the recombinant DNA technology, involving two main stages, the upstream and downstream stages. The first includes recombinant protein production in a suitable host microorganism, such as Escherichia coli (Castro et al, Sep. Purif. Technol., 2020), while the second comprises protein recovery, isolation, purification and polishing. Due to the high demands of the pharmaceutical industry for products with high purity and biological activity, the downstream stage is responsible for the majority of the production costs of biopharmaceuticals (50–90%), often including time-consuming and multi-step processes. Therefore, there is an immediate need to develop more efficient, cost-effective, and sustainable protein purification methodologies. In this work, two ionic-liquid-(IL)-based strategies were investigated for the purification of IFNα-2b recombinantly produced from E. coli fermentation broth, namely as adjuvants in aqueous biphasic systems or as chromatographic ligands immobilized in solid materials. Overall, the obtained results demonstrate that by tailoring IL’s chemical structures, improved protein purification processes are obtained and that the secondary structure of proteins is preserved.publishe

Sustainable ionic-liquid-based strategies for the downstream processing of interferon α-2b from Echerichia coli

Over the last decades, society has been facing an increment of chronic diseases due to the higher human life expectancy and the lack of efficient treatments for several pathologies. In this regard, biopharmaceuticals have become one of the most effective clinical treatments for a broad range of diseases, including cancer, metabolic and neurodegenerative disorders [1]. Among biopharmaceuticals, the role of interferons, particularly interferon α-2b (IFNα-2b), should be underlined, as they have been marketed for over 30 years with a considerable impact on the global therapeutic proteins market [2]. Usually based on the recombinant DNA technology, the manufacturing process of biopharmaceuticals encompasses two main stages: the upstream and downstream stages. Typically, the upstream phase includes recombinant protein production processes in a suitable host microorganism, such as Escherichia coli [3], while the general downstream processing of biopharmaceuticals comprises four stages - recovery, isolation, purification and polishing -, which are responsible for the majority of the production costs of biopharmaceuticals (50–90%) [3]. The downstream processing is a time-consuming and multi-step process, for which the development of cost-effective purification processes is mandatory to decrease their costs and environmental impact. In this context, two ionic-liquid-(IL)-based strategies were investigated in this work for the purification of IFNα-2b recombinantly produced from E. coli fermentation broth. ILs have been used as adjuvants in aqueous two-phase systems (ATPS) and applied in supported materials as alternative ligands. The obtained results demonstrate that ILs have a tailoring ability and contribute to the development of more effective and sustainable downstream processes of biopharmaceuticals.publishe

Differential expression of microRNAs in mouse pain models

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) are short non-coding RNAs that inhibit translation of target genes by binding to their mRNAs. The expression of numerous brain-specific miRNAs with a high degree of temporal and spatial specificity suggests that miRNAs play an important role in gene regulation in health and disease. Here we investigate the time course gene expression profile of miR-1, -16, and -206 in mouse dorsal root ganglion (DRG), and spinal cord dorsal horn under inflammatory and neuropathic pain conditions as well as following acute noxious stimulation.</p> <p>Results</p> <p>Quantitative real-time polymerase chain reaction analyses showed that the mature form of miR-1, -16 and -206, is expressed in DRG and the dorsal horn of the spinal cord. Moreover, CFA-induced inflammation significantly reduced miRs-1 and -16 expression in DRG whereas miR-206 was downregulated in a time dependent manner. Conversely, in the spinal dorsal horn all three miRNAs monitored were upregulated. After sciatic nerve partial ligation, miR-1 and -206 were downregulated in DRG with no change in the spinal dorsal horn. On the other hand, axotomy increases the relative expression of miR-1, -16, and 206 in a time-dependent fashion while in the dorsal horn there was a significant downregulation of miR-1. Acute noxious stimulation with capsaicin also increased the expression of miR-1 and -16 in DRG cells but, on the other hand, in the spinal dorsal horn only a high dose of capsaicin was able to downregulate miR-206 expression.</p> <p>Conclusions</p> <p>Our results indicate that miRNAs may participate in the regulatory mechanisms of genes associated with the pathophysiology of chronic pain as well as the nociceptive processing following acute noxious stimulation. We found substantial evidence that miRNAs are differentially regulated in DRG and the dorsal horn of the spinal cord under different pain states. Therefore, miRNA expression in the nociceptive system shows not only temporal and spatial specificity but is also stimulus-dependent.</p

Doença de Chagas no Brasil, foco no estado do Tocantins e a importância dos bancos de sangue para a identificação de novos casos

As the vector control of Triatoma infestans was conducted in Brazil, the transmission profile of Chagas disease (CD) has changed, with an increase of interest to other pathways, including transfusion. This study aimed to analyze the CD in Brazil, focusing on the state of Tocantins, and make an analysis of the importance of blood banks in the delimitation of new cases. Sectional study was conducted in a period from 2007 to 2014. The northern region, in which is situated the state of Tocantins, concentrates 90% of cases of Brazil. Today, 80% of the cities in Tocantins are at risk of having transmission of the disease due the invasion of homes by wild vector species, caused by the environmental impact resulted from the action of man in natural areas. The largest number of people infected in Tocantins is in the age group of potential blood donors. Due studies demonstrating the effectiveness in controlling transfusion transmission and methods of detection of CD in blood donors, we can conclude that blood banks have importance in the screening and diagnosis of new cases.Como el control contra el Triatoma infestans se llevó a cabo en Brasil, el perfil de la transmisión de la enfermedad de Chagas (CD) ha sido modificado con un aumento del interés de otras vías, incluyendo la transfusión. Pronto este estudio fue analizar el CD en Brasil, centrándose en el estado de Tocantins, y analizar la importancia de los bancos de sangre en la delimitación de los nuevos casos. Un estudio transversal se llevó a cabo en un período de 2007 a 2014. El Norte, que está en el estado de Tocantins, se concentra el 90% de los casos en Brasil. Hoy en día, el 80% de las ciudades de A están en riesgo de transmisión de enfermedades debido a la invasión de viviendas a la especie salvaje vector, causadas por el impacto ambiental de la actividad humana en áreas naturales. El mayor número de personas infectadas en Tocantins se encuentra en el grupo de edad de los potenciales donantes de sangre. Debido a los estudios que demuestran la eficacia en el control de transmisión y métodos de detección de CD en donantes de sangre de transfusión, podemos concluir que los bancos de sangre son importantes para la detección y el diagnóstico de nuevos casos.Como o controle do vetor Triatoma infestans foi realizada no Brasil, o perfil de transmissão da doença de Chagas (CD) foi modificado, com um aumento do interesse de outras vias, incluindo a transfusão. Logo este estudo teve como objetivo analisar o CD no Brasil, com foco no estado de Tocantins, e fazer uma análise da importância dos bancos de sangue na delimitação de novos casos. Um estudo transversal foi realizado em um período de 2007 a 2014. A região Norte, onde está situado o estado de Tocantins, concentra 90% dos casos do Brasil. Hoje em dia, 80% das cidades em TO estão em risco de ter a transmissão da doença, devido a invasão de casas por espécie selvagem vetor, causado pelo impacto ambiental resultante da ação do homem em áreas naturais. O maior número de pessoas infectadas no Tocantins está na faixa etária de potenciais doadores de sangue. Devido estudos que demonstram a eficácia no controle da transmissão e métodos de detecção de CD em doadores de sangue transfusão, podemos concluir que os bancos de sangue têm importância no rastreamento e diagnóstico de novos casos

Pesquisas na Universidade Federal do Tocantins e a Revista Desafios

A Universidade Federal do Tocantins&nbsp;(UFT)&nbsp;iniciou suas atividades em maio de 2003&nbsp;(PDI-UFT,&nbsp;2015)&nbsp;e a&nbsp;o longo&nbsp;destes anos foram criados cursos de&nbsp;graduação e pós-graduação nas diferentes áreas do conhecimento contando hoje com mais de 18.000 alunos, 1.029 professores e 854 técnicos-administrativos. Atualmente,&nbsp;a UFT está&nbsp;classificada&nbsp;no&nbsp;ranking&nbsp;do Guia do Estudante&nbsp;como uma das melhores Universidades da região norte (Guia do estudante, 2016).&nbsp;Concentrando&nbsp;mais de 85% do total&nbsp;dos&nbsp;doutores cadastrados na Plataforma Lattes&nbsp;no&nbsp;Estado do Tocantins&nbsp;que estão em atividade de&nbsp;Ensino,&nbsp;Pesquisa&nbsp;e Extensão&nbsp;(Plataforma Lattes,2016)&nbsp;o que&nbsp;conferea mesma, grande potencial&nbsp;para&nbsp;auxiliar o desenvolvimento científico e social do Estado. Conta com parcerias importantes tanto nacionais como internacionais

Biochemical quality of cowpea beans grown under water-deficit maintained with hydrogel

Investigations related to the effects of the hydrogel use in a water-deficit conditions on the biochemical composition of the harvested products are scarce. The aim of this study was to verify the influence of hydrogel on the biochemical response of cowpea beans produced under water-deficit in southern Tocantins. The experiment was a randomized complete block design with four replications. The treatments were obtained in a 2x5 factorial scheme, being: two sources of hydrogel (Hydroplan-EB HyC and Polim-Agri PP) and five concentrations (0, 5, 10, 15, and 20kg ha-1). After harvesting, the grains of cowpea, BRS Vinagre cultivar, were ground and chemically analyzed. The oil content of the seeds agreed with the reference values. The use of Polim-Agri PP promoted the best levels of amino acids (0.38mg g-1) and proteins (243.22mg g-1) of the grains. The carbohydrate content of them (Ȳ= 81.44mg g-1) was better using Hydroplan-EB HyC, regardless of the concentration applied. The application of 6.28kg ha-1 of Polim-Agri PP provided the highest starch content (842g mg-1). The use of hydrogel improves the biochemical quality of cowpea beans produced under water deficit in southern Tocantins

REMOÇÃO DE CÁLCIO DE EFLUENTE DE MÁQUINA DE PAPEL POR PRECIPITAÇÃO/COPRECIPITAÇÃO

In integrates pulp and paper mills, the effluent generated by the paper machine can be considered as a sector effluent, called white water, due to the high concentration of calcium. In this work, experiments were conducted to understand the behavior of the effluent in different pH values and to develop removal methods of calcium from the white water, aiming the reuse of water and the calcium recovery. Potentiometric titrations were carried out with HCl 0.022 mol L-1 and NaOH 0.025 mol L-1 standards, after adjusting the effluent pH at 12.0 and 2.0; respectively, which indicated inflection points for the carbonate, bicarbonate and kaolin, components capable of interaction with the soluble calcium. The methods for calcium removal consisted of coprecipitation/adsorption with iron (III) and aluminum hydroxides, and precipitation in the presence of sodium oxalate. The results indicated that at low concentrations of ferric sulfate and aluminum sulfate, the removal of calcium is low. In the adsorption assays in the presence of Fe(OH)3 and Al(OH)3, the increased of the ferric sulfate concentration enabled a slight increase in the calcium removal (16.5 to 31.0 %), reaching 65.0% in the adsorption more precipitation process in pH 10.0. In case of aluminum sulfate, the removal percentages were indifferent (close to 10.0%). In the precipitation of Ca2+ in the oxalate presence, the possibility of satisfactory percentages of removal was observed (75 to 87%), keeping the effluent with the conductivity and pH unchanged, it’s very important, because the increase of effluent conductivity to reuse cause break of paper made. The calcium oxalate recuperated can be heated excessively and changed and calcium carbonate and to be reused. Tests in the highest scale have to be realized to approve the reuse of water and calcium of paper machine.Nas fábricas integradas de celulose e papel, o efluente gerado pela máquina de papel pode ser considerado um efluente setorial, denominado de água branca, devido à concentração elevada de cálcio. Neste trabalho foram realizados experimentos com objetivos de compreender o comportamento do efluente em função do pH e desenvolver metodologias de remoção de cálcio da água branca, de modo a proporcionar o reuso da água e a recuperação de cálcio. Titulações potenciométricas foram realizadas com HCl 0,022 mol L-1 e com NaOH 0,025 mol L-1 padronizados, após ajuste de pH do efluente em 12,0 e 2,0 respectivamente, às quais indicaram pontos de inflexão referentes à carbonato, bicarbonato e caulim, componentes com capacidade de interação com o cálcio solúvel. Os métodos de remoção de cálcio consistiram de coprecipitação/adsorção com hidróxidos de ferro (III) e de alumínio e precipitação na presença de oxalato de sódio. Os resultados apontaram que em baixas concentrações de sulfato férrico e sulfato de alumínio, a coprecipitação de cálcio é baixa. Nos ensaios de adsorção na presença de Fe(OH)3 e Al(OH)3, notou-se que o aumento da concentração de sulfato férrico possibilitou um ligeiro aumento da remoção de cálcio (16,5 a 31,0%), chegando a 65,0% na soma dos processos de adsorção/precipitação em pH 10,0. No caso do sulfato de alumínio as porcentagens de remoção foram indiferentes (~ 10,0%). Em relação à precipitação de Ca2+ na presença de oxalato, observou-se remoção de porcentagens satisfatórias na forma de oxalato de cálcio (75% a 87%), mantendo-se o efluente com condutividade e pH praticamente inalterados, o que é muito desejável, já que o aumento da condutividade do efluente tratado na reutilização provoca efeito de quebra das folhas de papel. O oxalato de cálcio recuperado pode ser calcinado, sendo convertido a carbonato de cálcio, para posterior reuso. Testes em escala piloto devem ser realizados com o objetivo de validar o reuso da água e do cálcio provenientes da máquina de papel