RESUMO
La urgente necesidad de desarrollar y producir vacunas seguras y efectivas para garantizar la reducción de la propagación del coronavirus de tipo 2 causante del síndrome respiratorio agudo severo, hizo que el Centro de Inmunología Molecular y el Instituto Finlay de Vacunas, desarrollaran dos vacunas y un candidato vacunal contra la COVID-19, que tienen como componente la molécula del dominio de unión al receptor (aa 319-541) del virus. Para establecer el proceso productivo, se realizaron experimentos en los posibles pasos del proceso de purificación de la molécula del dominio de unión al receptor (aa 319-541), con vistas a su posterior transferencia tecnológica a escala industrial. Dicha molécula está fusionada con una etiqueta de hexahistidina en su extremo C-terminal y presenta nueve residuos de cisteína en su secuencia que forman cuatro enlaces disulfuros intramoleculares, quedando una cisteína libre que permite obtener dos moléculas: dimérica y monomérica, antígenos que forman parte de las vacunas SOBERANA®02 y SOBERANA®Plus y el candidato vacunal SOBERANA 01. Se determinaron las mejores condiciones de adsorción de las matrices cromatográficas de afinidad por quelatos metálicos, intercambio catiónico y exclusión molecular. Se evaluó el desempeño del proceso a escala piloto y se caracterizó la molécula de acuerdo a sus propiedades físico-químicas y biológicas. Los resultados obtenidos mostraron un 60,02 ± 5,15por ciento de recuperación total de la proteína de interés, con más del 98% de pureza en ambas moléculas, una eficiente remoción de contaminantes y una antigenicidad mayor del 90por ciento referido al monómero control del dominio de unión al receptor con 99 por ciento de pureza, lo que demuestra que el proceso establecido es eficiente en la obtención de un producto con la calidad requerida(AU)
The urgent need to develop and produce safe and effective vaccines to guarantee the reduction of the spread of the type 2 coronavirus that causes severe acute respiratory syndrome, led the Center for Molecular Immunology and the Finlay Vaccine Institute to develop two vaccines and one candidate vaccine to combat the 2019 coronavirus pandemic. As part of the establishment of the production process, experiments were carried out on the possible steps of the purification process of the receptor binding domain molecule (aa 319-541) with a view to its subsequent technological transfer on an industrial scale. This molecule is fused with a hexahistidine tag at its C-terminal end and has nine cysteine residues in its sequence that form four intramolecular disulfide bonds; leaving a free cysteine that allows two molecules to be obtained: dimeric and monomeric, which constitute the antigens of the SOBERANA®02 and SOBERANA®Plus vaccines and the SOBERANA 01 vaccine candidate. The best adsorption conditions of the chromatographic matrices of affinity for metal chelates, cationic exchange and molecular exclusion were determined. The performance of the process was evaluated on a pilot scale and the molecule was characterized according to its physical-chemical and biological properties. The results obtained showed a 60.02 ± 5.15percent total recovery of the protein of interest with more than 98% purity in both molecules, an efficient removal of contaminants and an antigenicity greater than 90percent referred to the control monomer of the domain receptor binding with 99% purity; which demonstrates that the established process is efficient in obtaining a product with the required quality(AU)
Assuntos
Humanos , Masculino , Feminino , Vacinas/imunologia , Vacinas contra COVID-19/uso terapêutico , Adsorção/efeitos dos fármacosRESUMO
Os resíduos provenientes da aquicultura são derivados da ração e da excreção dos peixes e podem estar sedimentados, suspensos ou dissolvidos, ocasionando elevados valores de DBO, DQO, nitrogênio e fósforo. A produção de camarões no Brasil tem gerado elevadas quantidades de resíduos sólidos, tendo em vista que os exoesqueletos dos camarões correspondem a cerca de 40% do seu peso total, resultando num forte impacto ambiental. Diversas pesquisas envolvendo a quitina estão sendo desenvolvidas na área de tratamento de água, devido principalmente a sua capacidade de formar filme, sendo utilizada em sistemas filtrantes. Este polissacarídeo também pode ser utilizado como agente floculante no tratamento de efluentes, como adsorvente na clarificação de óleos, e principalmente na produção de quitosana. Atualmente a quitosana possui aplicações multidimensionais, desde áreas como a nutrição humana, biotecnologia, ciência dos materiais, indústria farmacêutica, agricultura, terapia genética e proteção ambiental. A quitosana é muito eficiente na remoção de poluentes em diferentes concentrações. Apresenta alta capacidade e grande velocidade de adsorção, boa eficiência e seletividade tanto em soluções que possuem altas ou baixas concentrações. O uso da biotecnologia, através do processo de adsorção utilizando adsorventes naturais e baratos, como a quitina e quitosana, minimiza os impactos ambientais da aquicultura tanto em relação aos provocados pelo lançamento de efluentes no meio ambiente quanto aos causados pelo descarte inadequado dos resíduos do processamento de camarões.(AU)
Aquaculture residues are derived from fish feed and excretion and may be sedimented, suspended or dissolved, resulting in high BOD, COD, nitrogen and phosphorus values. Shrimp production in Brazil has generated high amounts of solid waste, since shrimp exoskeletons account for about 40% of their total weight, resulting in a strong environmental impact. Several researches involving chitin are being developed in the area of water treatment, mainly due to its ability to form film, being used in filter systems. This polysaccharide can also be used as a flocculating agent in the treatment of effluents, as an adsorbent in the clarification of oils, and especially in the production of chitosan. Currently, chitosan has multidimensional applications, from areas such as human nutrition, biotechnology, materials science, pharmaceutical industry, agriculture, gene therapy and environmental protection. Chitosan is very efficient in the removal of pollutants at different concentrations. It presents high capacity and high adsorption velocity, good efficiency and selectivity both in solutions that have high or low concentrations. The use of biotechnology, through the adsorption process using natural and cheap adsorbents such as chitin and chitosan, minimizes the environmental impacts of aquaculture both in relation to those caused by the release of effluents into the environment and those caused by the inappropriate disposal of processing residues of shrimps.(AU)
Los residuos procedentes de la acuicultura se derivan de la ración y de la excreción de los peces y pueden estar sedimentados, suspendidos o disueltos, ocasionando elevados valores de DBO, DQO, nitrógeno y fósforo. La producción de camarones en Brasil ha generado grandes cantidades de residuos sólidos, teniendo en cuenta que los exoesqueletos de los camarones corresponden a cerca del 40% de su peso total, resultando en un fuerte impacto ambiental. Varias investigaciones involucrando la quitina se están desarrollando en el área de tratamiento de agua, debido principalmente a su capacidad de formar película, siendo utilizada en sistemas filtrantes. Este polisacárido también puede ser utilizado como agente floculante en el tratamiento de efluentes, como adsorbente en la clarificación de aceites, y principalmente en la producción de quitosana. Actualmente la quitosana posee aplicaciones multidimensionales, desde áreas como la nutrición humana, biotecnología, ciencia de los materiales, industria farmacéutica, agricultura, terapia genética y protección ambiental. La quitosana es muy eficiente en la eliminación de contaminantes en diferentes concentraciones. Presenta alta capacidad y gran velocidad de adsorción, buena eficiencia y selectividad tanto en soluciones que poseen altas o bajas concentraciones. El uso de la biotecnología, a través del proceso de adsorción utilizando adsorbentes naturales y baratos, como la quitina y quitosana, minimiza los impactos ambientales de la acuicultura tanto en relación a los provocados por el lanzamiento de efluentes en el medio ambiente en cuanto a los causados por el descarte inadecuado de los residuos del procesamiento de camarones.(AU)
Assuntos
Quitina/administração & dosagem , Adsorção/efeitos dos fármacos , Quitosana/administração & dosagem , Águas Residuárias/química , Biopolímeros/análise , Aquicultura , Eutrofização/fisiologia , Amônia/químicaRESUMO
One of the hallmarks of Alzheimer's Disease (AD) is the anomalous binding involving amyloid-ß (Aß) peptide and metal ions, such as copper, formed through histidine (His) residues. Herein, adsorption experiments were performed to test the in vitro ability of chitosan to uptake copper ions in the presence of histidine. The characterization of the beads was assessed before and after the adsorption process by scanning electron microscope, X-ray diffraction and Fourier-transform infrared spectroscopy. Amino acid functionalization of chitosan-based beads promoted an increase in the copper ions adsorption capacity (2.47 mmol of Cu(II)/gram of adsorbent). Nevertheless, depending on the order of addition of histidine to the system, different adsorption behaviors were observed. The kinetics showed that, once the Cu(II)-His bond was established, functionalized beads were less efficient to capture Cu(II), which promoted a decrease in the overall adsorption capacity. However, when chitosan and histidine were simultaneously added to the Cu(II) solution, there was no decrease in adsorption capacity. To sum up, chitosan-based materials are an interesting model to provide a better understanding on the biomoleculescopper interactions that occur in AD, as well as a possible chelating agent that can interfere in the bonds between Aß residues and copper ions.
Assuntos
Peptídeos beta-Amiloides/química , Quitosana/química , Cobre/química , Histidina/química , Adsorção/efeitos dos fármacos , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Aminoácidos/química , Quitosana/farmacologia , Humanos , Íons/química , Cinética , Microscopia Eletrônica de Varredura , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
In this research, the adsorption of three synthetic dyes dissolved in an aqueous solution on chitosan cryogel beads (Q-C-EGDE) was compared. The effect of the pH of the solution on the adsorption capacity of each dyes was analyzed. Furthermore, the kinetics and adsorption isotherms were compared, at temperatures of 283.15 K, 303.15 K and 323.15 K, and the kinetic and adsorption equilibrium data were fitted to three mathematical models, respectively. The biosorbent was characterized by scanning electron microscopy (SEM), the nitrogen physisorption BET method and Fourier transform infrared spectroscopy (FTIR). The characterization results show that the cryogel is composed of low-surface, macroporous, porous grooved walls. The functional groups that took part in the adsorption were mainly amino groups (NH3+). When comparing the adsorption capacities, it was found that the dyes adsorb in the following order Blue 1 > Red 2 > Yellow 5 reaching capacities from 1600 mg/L to 850 mg/L. The results of the adsorption and mathematical modelling suggest that the process is regulated mainly by physisorption and is largely limited by mass transfer mechanisms within the cryogel, where the electrostatic charges present affect adsorption. The latter was corroborated by the Monte Carlo simulation.
Assuntos
Quitosana/química , Corantes/química , Criogéis/química , Adsorção/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Cinética , Microscopia Eletrônica de Varredura/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Temperatura , Poluentes Químicos da Água/química , Purificação da Água/métodosRESUMO
Films made from neat chitosan and chitosan with magnetic nanoparticles (MNPs) were tested as adsorbents of arsenate ions. Sorption equilibrium and sorption kinetics studies are reported, including different models applied to enlighten experimental observations and predict results. The sorption of As (V) was reasonably explained using Freundlich isotherm for neat chitosan film although it was better represented by Langmuir equation for the composite sample. The experimental kinetics results showed that the adsorption of arsenate ions is very fast during the first minutes and then the composite seems to reach saturation, while a slow desorption in the chitosan film was observed and acceptably fitted with a pseudo first order reversible model. The adsorbent containing MNPs presented higher adsorption capacity, which was associated to the additional adsorbent capacity provided by the MNPs and its much more irregular surface area that leads to an enhanced adsorption surface. For instance, at 10 mg/L equilibrium concentration, which corresponds to an initial concentration of As (V) much higher than the normal concentration of arsenate in natural water, chitosan-MNP sample exhibits a removal capacity of 10.4 mg/g that is more than six times higher than the 1.6 mg/g shown by the chitosan film.
Assuntos
Arsênio/isolamento & purificação , Quitosana/química , Poluentes Químicos da Água/isolamento & purificação , Purificação da Água/métodos , Adsorção/efeitos dos fármacos , Arsênio/toxicidade , Compostos Férricos/química , Cinética , Nanopartículas/química , Água/química , Poluentes Químicos da Água/toxicidadeRESUMO
: In this study, the mechanisms of methylparaben adsorption onto activated carbon (AC) are elucidated starting from equilibrium and thermodynamic data. Adsorption tests are carried out on three ACs with different surface chemistry, in different pH and ionic strength aqueous solutions. Experimental results show that the methylparaben adsorption capacity is slightly affected by pH changes, while it is significantly reduced in the presence of high ionic strength. In particular, methylparaben adsorption is directly dependent on the micropore volume of the ACs and the π- stacking interactions, the latter representing the main interaction mechanism of methylparaben adsorption from liquid phase. The equilibrium adsorption data are complemented with novel calorimetric data that allow calculation of the enthalpy change associated with the interactions between solvent-adsorbent, adsorbent-adsorbate and the contribution of the ester functional group (in the methylparaben structure) to the adsorbateâ»adsorbent interactions, in different pH and ionic strength conditions. It was determined that the interaction enthalpy of methylparaben-AC in water increases (absolute value) slightly with the basicity of the activated carbons, due to the formation of interactions with π- electrons and basic functional groups of ACs. The contribution of the ester group to the adsorbate-adsorbent interactions occurs only in the presence of phenol groups on AC by the formation of Brønstedâ»Lowry acidâ»base interactions.
Assuntos
Adsorção/efeitos dos fármacos , Parabenos/química , Termodinâmica , Poluentes Químicos da Água/química , Calorimetria , Carbono/química , Carvão Vegetal/química , Fenóis/química , Solventes/química , Água/química , Poluentes Químicos da Água/toxicidadeRESUMO
This study investigated the effect of surfactants associated with sodium fluoride (NaF) on enamel erosion prevention, using an erosion-remineralization in vitro model. Sodium lauryl sulfate (SLS), polysorbate 20 (P20), and cocoamidopropyl betaine (CAPB) were tested, at concentrations of 1.0 and 1.5%, and associated or not with NaF (275 ppm). The control groups were distilled water and the NaF solution. Bovine enamel samples (n = 12) were prepared and submitted to a 5-day cycling model: acid challenge (0.3% citric acid, pH 2.6, 4×/day), human saliva (2 h, 4×/day), and the treatment solutions (2 min, 2×/day). The protective potential of the agents against initial erosion was assessed by microhardness and the surface loss by profilometry. Enamel surface wettability was determined by goniometry, protein adsorption was measured by spectroscopy (FTIR), and the KOH-soluble fluoride was quantified. Goniometry showed that SLS and CAPB increased enamel wettability. No differences were found among the surfactants regarding protein adsorption. Microhardness showed that SLS reduced NaF protection. P20 (1 and 1.5%) and CAPB 1.5% presented a protective effect, but lower than the NaF solution. Profilometry showed that CAPB protected enamel, but no agent associated with NaF promoted a higher protection than the NaF solution alone. KOH-soluble fluoride analysis showed that all surfactants reduced the fluoride adsorption on the enamel surface. Therefore, the surfactants tested (except for P20) changed the enamel surface energy. The SLS decreased the protective potential of NaF on initial erosion, but no tested agent interfered with the protective effect of NaF on enamel erosive wear.
Assuntos
Betaína/análogos & derivados , Cariostáticos/farmacologia , Esmalte Dentário/patologia , Polissorbatos/farmacologia , Dodecilsulfato de Sódio/farmacologia , Fluoreto de Sódio/farmacologia , Tensoativos/farmacologia , Erosão Dentária/prevenção & controle , Adsorção/efeitos dos fármacos , Análise de Variância , Animais , Betaína/química , Betaína/farmacologia , Bovinos , Ácido Cítrico/efeitos adversos , Esmalte Dentário/efeitos dos fármacos , Dureza , Polissorbatos/química , Saliva/fisiologia , Dodecilsulfato de Sódio/química , Tensoativos/química , Erosão Dentária/induzido quimicamente , Molhabilidade/efeitos dos fármacos , Espectroscopia por Absorção de Raios XRESUMO
AIM: To investigate the effects of combinations of several irrigants on the roughness and wettability of dentine, adhesion of Enterococcus faecalis and Candida albicans and adsorption of chlorhexidine (CHX) to the dentine. METHODOLOGY: Bovine dentine samples were prepared and their surface roughness standardized. The samples were distributed in groups (n = 10) and subjected to one of the following irrigation protocols: G1 - saline solution; G2 - sodium hypochlorite (NaOCl); G3 - NaOCl + ethylenediaminetetraacetic acid (EDTA); G4 - NaOCl + peracetic acid (PAA); G5 - NaOCl + 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP); G6 - NaOCl + EDTA + CHX; G7 - NaOCl + PAA + CHX; G8 - NaOCl + HEDP + CHX; and G9 - mixture of NaOCl + HEDP. After treatments, roughness and wettability were measured. In order to evaluate the adhesion of microorganisms to dentine, new dentine samples were prepared and after 2 h of contact with the microorganisms, were analysed using a confocal laser scanning microscope and the number of microorganisms adhering to the surfaces were determined. Absorption spectra were collected by attenuated total reflectance of Fourier-transform infrared spectroscopy before and after immersion of other dentine samples in each solution of G6, G7 and G8 and in a solution of 2% CHX at various time intervals. The areas of the band associated with CHX with the peak at 1492 cm-1 were calculated between 1479 and 1500 cm-1 of the spectral range. The data obtained in all experiments were subjected to one-way ANOVA (α < 0.05). The values of the CHX band were also subjected to one-way repeated measures ANOVA (α < 0.05). RESULTS: Saline solution, NaOCl, HEDP and CHX did not alter the roughness of the dentine (P > 0.05), whilst EDTA and PAA did (P < 0.05). Dentine surface wettability increased after the use of all irrigants compared to saline solution (P < 0.05), with HEDP causing the greatest increases (P < 0.05). The adhesion of E. faecalis was favoured on surfaces treated with only saline solution and NaOCl, and on samples that had decalcifying agents as the final irrigant (P < 0.05). The adhesion of C. albicans was highest on surfaces treated with only saline solution and on surfaces that had NaOCl used as the last irrigant (P < 0.05). The use of CHX as a final irrigant reduced the adhesion of both microorganisms. The roughness and wettability did not influence the adhesion of the microorganisms tested. The adsorption of CHX to the dentine was significant after 1 min of immersion of the mineralized samples in the irrigant (P < 0.05), and the use of chelating agents prior to CHX potentiated this adsorption. CONCLUSIONS: The irrigation solutions had a variable effect on the properties of dentine, on the adhesion of E. faecalis and C. albicans and the adsorption of CHX to the dentine surface.
Assuntos
Aderência Bacteriana/efeitos dos fármacos , Clorexidina/farmacologia , Dentina/efeitos dos fármacos , Dentina/microbiologia , Dentina/patologia , Irrigantes do Canal Radicular/farmacologia , Adsorção/efeitos dos fármacos , Animais , Candida albicans/efeitos dos fármacos , Bovinos , Quelantes/farmacologia , Ácido Edético/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Ácido Etidrônico/farmacologia , Teste de Materiais , Ácido Peracético/farmacologia , Hipoclorito de Sódio/farmacologia , Propriedades de Superfície/efeitos dos fármacosRESUMO
Chitosan partially functionalized with aldehyde groups was used for enzyme immobilization, favoring first the enzyme adsorption through its amino groups and then the covalent bonding of the adsorbed catalyst through the aldehyde groups of the support. Using this strategy, immobilized A. oryzae ß-galactosidase had a better performance than when only the aldehyde groups were used. The performance was further improved by modifying the support aldehyde group density to 200⯵molesâ g-1. The biocatalyst under optimized immobilization conditions had 2951â¯IUâ g-1 and half-life of 46.3â¯min at 60⯰C, while its agarose counterpart had 2294â¯IUâ g-1 and half-life of 59.5â¯min. Both biocatalysts were applied in galacto-oligosaccharide synthesis. After 10 sequential batches, the cumulative productivity (gGOSâ h-1Ëgprotein-1) obtained with the chitosan and the agarose biocatalysts were 4.7 and 4.0 times the value when soluble enzyme was used respectively. This methodology had not been reported previously with chitosan, showing the high versatility of this low cost carrier and its high potential for enzyme immobilization.
Assuntos
Quitosana/metabolismo , Enzimas Imobilizadas/metabolismo , beta-Galactosidase/metabolismo , Adsorção/efeitos dos fármacos , Aldeídos/metabolismo , Catálise/efeitos dos fármacos , Sefarose/metabolismoRESUMO
Zinc and cadmium can be efficiently removed from solutions using the brown algae, Macrocystis pyrifera. Treatment with CaCl(2) allowed stabilization of the biosorbent. The maximum biosorption capacities in mono-component systems were 0.91 mmol g(-1) and 0.89 mmol g(-1) and the Langmuir affinity coefficients were 1.76 L mmol(-1) and 1.25 L mmol(-1) for Zn(II) and Cd(II), respectively. In two-component systems, Zn(II) and Cd(II) adsorption capacities were reduced by 50% and 40%, respectively and the biosorbent showed a preference for Cd(II) over Zn(II). HNO(3) (0.1M) and EDTA (0.1M) achieved 90-100% desorption of both ions from the loaded biomass. While HNO(3) preserved the biomass structure, EDTA destroyed it completely. Fourier transform infrared spectra identified the contribution of carboxylic, amine and sulfonate groups on Zn(II) and Cd(II) biosorption. These results showed that biosorption using M. pyrifera-treated biomass could be an affordable and simple process for cadmium and zinc removal from wastewaters.