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Mara G. Freire graduated in Chemistry in 2003 by University of Aveiro, Portugal, receiving the “Best Chemistry Student Award” from Dow Portugal. By the end of 2007 she completed her PhD in Chemical Engineering by University of Aveiro, with trainees at Federal University of Rio de Janeiro, Brazil, and Claude Bernard University, Lyon, France, followed by post-doctoral activities at ITQB2, New University of Lisbon, Portugal. In 2012, Freire was an invited professor at Tiradentes University, Sergipe, Brazil. From June 2013 to January 2014, Freire was an Assistant Researcher at CICECO - Aveiro Institute of Materials, Chemistry Department, University of Aveiro, Portugal, and since February 2014 she is a Coordinator Researcher at the same institution. Currently, Freire is the Coordinator of Group 5 of CICECO, Biomedical and Biomimetic Materials, and Deputy Chair of the Scientific Council of University of Aveiro. Freire published +250 papers in international peer reviewed journals, +20 book chapters and edited one book, and has +14,000 citations and h-index of 67. She coordinated/participated in +20 R&D projects, among which a Starting and a Proof of Concept Grants from the European Research Council (ERC). She has filled +10 national/international patents, and is the co-founder of the RYAPURTECH spin-off. Up to date, she has completed the supervision of 18 post-doctoral researchers, and 14 PhD, 53 MSc and 40 BSc students.
L-asparaginase (ASNase) is an aminohydrolase enzyme widely used in the pharmaceutical and food industries. Although currently its main applications are focused on the treatment of lymphoproliferative disorders such as acute lymphoblastic leukemia (ALL) and acrylamide reduction in starch-rich foods cooked at temperatures above 100 °C, its use as a biosensor in the detection and monitoring of L-asparagine levels is of high relevance. ASNase-based biosensors are a promising and innovative technology, mostly based on colorimetric detection since the mechanism of action of ASNase is the catalysis of the L-asparagine hydrolysis, which releases L-aspartic acid and ammonium ions, promoting a medium pH value change followed by color variation. ASNase biosensing systems prove their potential for L-asparagine monitoring in ALL patients, along with L-asparagine concentration analysis in foods, due to their simplicity and fast response.
João C. F. Nunes; Raquel O. Cristóvão; Valéria C. Santos-Ebinuma; Joaquim L. Faria; Cláudia G. Silva; Márcia C. Neves; Mara G. Freire; Ana P. M. Tavares. L-Asparaginase-Based Biosensors. Encyclopedia 2021, 1, 848 -858.
AMA StyleJoão C. F. Nunes, Raquel O. Cristóvão, Valéria C. Santos-Ebinuma, Joaquim L. Faria, Cláudia G. Silva, Márcia C. Neves, Mara G. Freire, Ana P. M. Tavares. L-Asparaginase-Based Biosensors. Encyclopedia. 2021; 1 (3):848-858.
Chicago/Turabian StyleJoão C. F. Nunes; Raquel O. Cristóvão; Valéria C. Santos-Ebinuma; Joaquim L. Faria; Cláudia G. Silva; Márcia C. Neves; Mara G. Freire; Ana P. M. Tavares. 2021. "L-Asparaginase-Based Biosensors." Encyclopedia 1, no. 3: 848-858.
Rutin is a known antioxidant compound that displays a broad range of biological activities and health-related benefits but presents a low water solubility that can be overcome by its polymerization. In this work, biocompatible aqueous biphasic systems composed of the ionic liquid cholinium dihydrogen phosphate ([CH][DHph]) and the polymer poly(ethylene glycol) 600 (PEG 600) were investigated as an efficient integrated reaction–separation platform for the laccase-catalyzed oligomerization of rutin. Two different approaches were studied to reuse laccase in several oligorutin production cycles, the main difference between them being the use of monophasic or biphasic regimes during the oligomerization reaction. The use of a biphasic regime in the second approach (heterogeneous reaction medium) allowed the successful reuse of the biocatalyst in three consecutive reaction–separation cycles while achieving noteworthy rutin oligomerization yields (95% in the first cycle, 91% in the second cycle, and 89% in the last cycle). These remarkable results were caused by the combination of the increased solubility of rutin in the PEG-rich phase together with the enhanced catalytic performance of laccase in the [Ch][DHph]-rich phase, alongside with the optimization of the pH of the reaction medium straightly linked to enzyme stability. Finally, a life-cycle assessment was performed to compare this integrated reaction–separation platform to three alternative processes, reinforcing its sustainability.
Abel Muñiz-Mouro; Ana M. Ferreira; João A. P. Coutinho; Mara G. Freire; Ana P. M. Tavares; Patricia Gullón; Sara González-García; Gemma Eibes. Integrated Biocatalytic Platform Based on Aqueous Biphasic Systems for the Sustainable Oligomerization of Rutin. ACS Sustainable Chemistry & Engineering 2021, 9, 9941 -9950.
AMA StyleAbel Muñiz-Mouro, Ana M. Ferreira, João A. P. Coutinho, Mara G. Freire, Ana P. M. Tavares, Patricia Gullón, Sara González-García, Gemma Eibes. Integrated Biocatalytic Platform Based on Aqueous Biphasic Systems for the Sustainable Oligomerization of Rutin. ACS Sustainable Chemistry & Engineering. 2021; 9 (29):9941-9950.
Chicago/Turabian StyleAbel Muñiz-Mouro; Ana M. Ferreira; João A. P. Coutinho; Mara G. Freire; Ana P. M. Tavares; Patricia Gullón; Sara González-García; Gemma Eibes. 2021. "Integrated Biocatalytic Platform Based on Aqueous Biphasic Systems for the Sustainable Oligomerization of Rutin." ACS Sustainable Chemistry & Engineering 9, no. 29: 9941-9950.
Spent coffee grounds (SCGs) are a waste product with no relevant commercial value. However, SCGs are rich in extractable compounds with biological activity. To add value to this coffee byproduct, water and aqueous solutions of cholinium-based ionic liquids (ILs) were studied to extract caffeine from SCGs. In general, all IL aqueous solutions lead to higher extraction efficiencies of caffeine than pure water, with aqueous solutions of cholinium bicarbonate being the most efficient. A factorial planning was applied to optimize operational conditions. Aqueous solutions of cholinium bicarbonate, at a temperature of 80 °C for 30 min of extraction, a biomass–solvent weight ratio of 0.05 and at an IL concentration of 1.5 M, made it possible to extract 3.29 wt% of caffeine (against 1.50 wt% obtained at the best conditions obtained with pure water). Furthermore, to improve the sustainability of the process, the same IL aqueous solution was consecutively applied to extract caffeine from six samples of fresh biomass, where an increase in the extraction yield from 3.29 to 13.10 wt% was achieved. Finally, the cholinium bicarbonate was converted to cholinium chloride by titration with hydrochloric acid envisioning the direct application of the IL-caffeine extract in food, cosmetic and nutraceutical products. The results obtained prove that aqueous solutions of cholinium-based ILs are improved solvents for the extraction of caffeine from SCGs, paving the way for their use in the valorization of other waste rich in high-value compounds.
Ana Ferreira; Hugo Gomes; João Coutinho; Mara Freire. Valorization of Spent Coffee by Caffeine Extraction Using Aqueous Solutions of Cholinium-Based Ionic Liquids. Sustainability 2021, 13, 7509 .
AMA StyleAna Ferreira, Hugo Gomes, João Coutinho, Mara Freire. Valorization of Spent Coffee by Caffeine Extraction Using Aqueous Solutions of Cholinium-Based Ionic Liquids. Sustainability. 2021; 13 (13):7509.
Chicago/Turabian StyleAna Ferreira; Hugo Gomes; João Coutinho; Mara Freire. 2021. "Valorization of Spent Coffee by Caffeine Extraction Using Aqueous Solutions of Cholinium-Based Ionic Liquids." Sustainability 13, no. 13: 7509.
Green Chemistry has been defined by the EPA as the design of chemical products and processes that reduce or eliminate the use or generation of hazardous substances
Gonzalo de Gonzalo; Mara Freire. 25th Anniversary of Molecules—Recent Advances in Green Chemistry. Molecules 2021, 26, 3768 .
AMA StyleGonzalo de Gonzalo, Mara Freire. 25th Anniversary of Molecules—Recent Advances in Green Chemistry. Molecules. 2021; 26 (12):3768.
Chicago/Turabian StyleGonzalo de Gonzalo; Mara Freire. 2021. "25th Anniversary of Molecules—Recent Advances in Green Chemistry." Molecules 26, no. 12: 3768.
In the past decades, the production of biopharmaceuticals has gained high interest due to its great sensitivity, specificity, and lower risk of negative effects to patients. Biopharmaceuticals are mostly therapeutic recombinant proteins produced through biotechnological processes. In this context, L-asparaginase (L-asparagine amidohydrolase, L-ASNase (E.C. 3.5.1.1)) is a therapeutic enzyme that has been abundantly studied by researchers due to its antineoplastic properties. As a biopharmaceutical, L-ASNase has been used in the treatment of acute lymphoblastic leukemia (ALL), acute myeloblastic leukemia (AML), and other lymphoid malignancies, in combination with other drugs. Besides its application as a biopharmaceutical, this enzyme is widely used in food processing industries as an acrylamide mitigation agent and as a biosensor for the detection of L-asparagine in physiological fluids at nano-levels. The great demand for L-ASNase is supplied by recombinant enzymes from Escherichia coli and Erwinia chrysanthemi. However, production processes are associated to low yields and proteins associated to immunogenicity problems, which leads to the search for a better enzyme source. Considering the L-ASNase pharmacological and food importance, this review provides an overview of the current biotechnological developments in L-ASNase production and biochemical characterization aiming to improve the knowledge about its production. • Microbial enzyme applications as biopharmaceutical and in food industry • Biosynthesis process: from the microorganism to bioreactor technology • Enzyme activity and kinetic properties: crucial for the final application
Daniel Castro; Ana Sofia C. Marques; Mafalda R. Almeida; Gabriela B. de Paiva; Heitor B. S. Bento; Danielle B. Pedrolli; Mara G. Freire; Ana P. M. Tavares; Valéria C. Santos-Ebinuma. L-asparaginase production review: bioprocess design and biochemical characteristics. Applied Microbiology and Biotechnology 2021, 105, 4515 -4534.
AMA StyleDaniel Castro, Ana Sofia C. Marques, Mafalda R. Almeida, Gabriela B. de Paiva, Heitor B. S. Bento, Danielle B. Pedrolli, Mara G. Freire, Ana P. M. Tavares, Valéria C. Santos-Ebinuma. L-asparaginase production review: bioprocess design and biochemical characteristics. Applied Microbiology and Biotechnology. 2021; 105 (11):4515-4534.
Chicago/Turabian StyleDaniel Castro; Ana Sofia C. Marques; Mafalda R. Almeida; Gabriela B. de Paiva; Heitor B. S. Bento; Danielle B. Pedrolli; Mara G. Freire; Ana P. M. Tavares; Valéria C. Santos-Ebinuma. 2021. "L-asparaginase production review: bioprocess design and biochemical characteristics." Applied Microbiology and Biotechnology 105, no. 11: 4515-4534.
Egg white proteins (ovalbumin, ovotransferrin, lysozyme, ovoinhibitor and ovomucin) are used in a wide range of applications, thus requiring the development of cost-effective purification platforms for their isolation and recovery. In this work, aqueous biphasic systems (ABS) and three phase partitioning (TPP) were investigated as combined approaches to separate egg white proteins. ABS composed of poly(ethylene)glycol of different molecular weights and potassium phosphate buffer at pH 7 were first characterized and optimized to simultaneously separate ovalbumin and lysozyme. Within the ABS concept, there is the preferential partition of both proteins to the PEG-rich phase. However, by increasing the PEG molecular weight or phase-forming components content, there is the selective precipitation of lysozyme over ovalbumin at the ABS interface, falling within the TPP concept. The best TPP systems were then applied to separate proteins directly from egg white. With the best TPP identified, there is the fractionation of 3 main proteins from egg yolk, namely ovalbumin in the PEG-rich phase, lysozyme at the ABS interface and ovoinhibitor in the salt-rich phase. The obtained results reinforce the potential of ABS/TTP combined strategies to fractionate high-value proteins from complex matrices in a single-step.
Diana C.V. Belchior; Mara G. Freire. Simultaneous separation of egg white proteins using aqueous three-phase partitioning systems. Journal of Molecular Liquids 2021, 336, 116245 .
AMA StyleDiana C.V. Belchior, Mara G. Freire. Simultaneous separation of egg white proteins using aqueous three-phase partitioning systems. Journal of Molecular Liquids. 2021; 336 ():116245.
Chicago/Turabian StyleDiana C.V. Belchior; Mara G. Freire. 2021. "Simultaneous separation of egg white proteins using aqueous three-phase partitioning systems." Journal of Molecular Liquids 336, no. : 116245.
Proteins are one the most widely studied biomolecules with diverse functions and applications. Aiming at overcoming the current drawbacks of purification processes of proteins, the introduction of ionic liquids (ILs) has been a hot topic of research. ILs have been applied in the creation of aqueous biphasic systems (IL-based ABS), solid-phase extractions through poly(ionic liquid)s (PILs) and supported ionic-liquid phases (SILPs), and in the crystallization of proteins. In this sense, ILs have emerged as solvents, electrolytes or adjuvants, or as supported materials to tune the adsorption/affinity capacity aiming at developing an efficient, cost-effective, sustainable and green IL-based process for protein extraction. This review discusses different IL-based processes in the extraction and purification of proteins in the past years, namely IL-based aqueous biphasic systems (IL-based ABS), solid-phase extractions through PILs and SILPs, and protein crystallization. The type and structure of ILs applied and their influence in the different processes performance are also discussed.
João C. F. Nunes; Mafalda R. Almeida; Joaquim L. Faria; Cláudia G. Silva; Márcia C. Neves; Mara G. Freire; Ana P. M. Tavares. Overview on Protein Extraction and Purification Using Ionic-Liquid-Based Processes. Journal of Solution Chemistry 2021, 1 -36.
AMA StyleJoão C. F. Nunes, Mafalda R. Almeida, Joaquim L. Faria, Cláudia G. Silva, Márcia C. Neves, Mara G. Freire, Ana P. M. Tavares. Overview on Protein Extraction and Purification Using Ionic-Liquid-Based Processes. Journal of Solution Chemistry. 2021; ():1-36.
Chicago/Turabian StyleJoão C. F. Nunes; Mafalda R. Almeida; Joaquim L. Faria; Cláudia G. Silva; Márcia C. Neves; Mara G. Freire; Ana P. M. Tavares. 2021. "Overview on Protein Extraction and Purification Using Ionic-Liquid-Based Processes." Journal of Solution Chemistry , no. : 1-36.
The advent of biopharmaceuticals in modern medicine brought enormous benefits to the treatment of numerous human diseases and improved the well-being of many people worldwide. First introduced in the market in the early 1980s, the number of approved biopharmaceutical products has been steadily increasing, with therapeutic proteins, antibodies, and their derivatives accounting for most of the generated revenues. The success of pharmaceutical biotechnology is closely linked with remarkable developments in DNA recombinant technology, which has enabled the production of proteins with high specificity. Among promising biopharmaceuticals are interferons, first described by Isaacs and Lindenmann in 1957 and approved for clinical use in humans nearly thirty years later. Interferons are secreted autocrine and paracrine proteins, which by regulating several biochemical pathways have a spectrum of clinical effectiveness against viral infections, malignant diseases, and multiple sclerosis. Given their relevance and sustained market share, this review provides an overview on the evolution of interferon manufacture, comprising their production, purification, and formulation stages. Remarkable developments achieved in the last decades are herein discussed in three main sections: (i) an upstream stage, including genetically engineered genes, vectors, and hosts, and optimization of culture conditions (culture media, induction temperature, type and concentration of inducer, induction regimens, and scale); (ii) a downstream stage, focusing on single- and multiple-step chromatography, and emerging alternatives (e.g., aqueous two-phase systems); and (iii) formulation and delivery, providing an overview of improved bioactivities and extended half-lives and targeted delivery to the site of action. This review ends with an outlook and foreseeable prospects for underdeveloped aspects of biopharma research involving human interferons.
Leonor Castro; Guilherme Lobo; Patrícia Pereira; Mara Freire; Márcia Neves; Augusto Pedro. Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation. Vaccines 2021, 9, 328 .
AMA StyleLeonor Castro, Guilherme Lobo, Patrícia Pereira, Mara Freire, Márcia Neves, Augusto Pedro. Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation. Vaccines. 2021; 9 (4):328.
Chicago/Turabian StyleLeonor Castro; Guilherme Lobo; Patrícia Pereira; Mara Freire; Márcia Neves; Augusto Pedro. 2021. "Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation." Vaccines 9, no. 4: 328.
Ionic liquids (ILs) are molten salts composed of a large organic cation and an organic/inorganic anion. Due to their ionic character, most ILs present advantageous properties over conventional solvents, such as negligible volatility at atmospheric conditions and high thermal and chemical stabilities. The wide variety of IL anion–cation combinations allows these solvents to be designed to display a strong solvation ability for a myriad of active pharmaceutical ingredients (APIs) and (bio)polymers. Given these properties, ILs have been used as solvents and as formulation components in different areas of drug delivery, as well as novel liquid forms of APIs (API-ILs) applied in different stages of development of novel drug delivery systems. Furthermore, their combination with polymers and biopolymers has enabled the design of drug delivery systems for new therapeutic routes of administration.
Sónia Pedro; Carmen Freire; Armando Silvestre; Mara Freire. Ionic Liquids in Drug Delivery. Encyclopedia 2021, 1, 324 -339.
AMA StyleSónia Pedro, Carmen Freire, Armando Silvestre, Mara Freire. Ionic Liquids in Drug Delivery. Encyclopedia. 2021; 1 (2):324-339.
Chicago/Turabian StyleSónia Pedro; Carmen Freire; Armando Silvestre; Mara Freire. 2021. "Ionic Liquids in Drug Delivery." Encyclopedia 1, no. 2: 324-339.
Novel liquid supports for enzyme immobilization and reuse based on aqueous biphasic systems (ABS) constituted by cholinium‐based ionic liquids (ILs) and polymers for the degradation of dyes are here proposed. The biocatalytic reaction for dye decolorization using laccase occurs in the biphasic medium, with the enzyme being “supported” in the IL‐rich phase and the dye and degradation products being enriched in the polymer‐rich phase. An initial screening of the laccase activity in aqueous solutions of ABS constituents, namely cholinium dihydrogen citrate ([Ch][DHC]), cholinium dihydrogen phosphate ([Ch][DHP]), cholinium acetate ([Ch][Acet]), polypropylene glycol 400 (PPG 400), polyethylene glycol 400 (PEG 400) and K2HPO4 was carried out. Compared to the buffered control, a relative laccase activity of up to 170%, 257% and 530% was observed with PEG 400, [Ch][DHP] and [Ch][DHC], respectively. These ABS constituents were then investigated for the in situ enzymatic biodegradation of the Remazol Brilliant Blue R (RBBR) dye. At the optimized conditions, the ABS constituted PPG 400 at 46 wt% and [Ch][DHC] at 16 wt% leads to the complete degradation of the RBBR dye, further maintaining the enzyme activity. This ABS also allows an easy immobilization, recovery and reuse of the biocatalyst for six consecutive reaction cycles, achieving a degradation yield of the dye of 96% in the last cycle. In summary, if properly designed, high enzymatic activities and reaction yields are obtained with ABS as liquid supports, while simultaneously overcoming the safety and environmental concerns of conventional organic solvents used in liquid‐liquid heterogeneous reactions, thus representing more sustainable biocatalytic processes.
Ana M. Ferreira; Ana I. Valente; Leonor S. Castro; João A. P. Coutinho; Mara G. Freire; Ana P. M. Tavares. Sustainable liquid supports for laccase immobilization and reuse: Degradation of dyes in aqueous biphasic systems. Biotechnology and Bioengineering 2021, 118, 2514 -2523.
AMA StyleAna M. Ferreira, Ana I. Valente, Leonor S. Castro, João A. P. Coutinho, Mara G. Freire, Ana P. M. Tavares. Sustainable liquid supports for laccase immobilization and reuse: Degradation of dyes in aqueous biphasic systems. Biotechnology and Bioengineering. 2021; 118 (7):2514-2523.
Chicago/Turabian StyleAna M. Ferreira; Ana I. Valente; Leonor S. Castro; João A. P. Coutinho; Mara G. Freire; Ana P. M. Tavares. 2021. "Sustainable liquid supports for laccase immobilization and reuse: Degradation of dyes in aqueous biphasic systems." Biotechnology and Bioengineering 118, no. 7: 2514-2523.
Polymer–polymer aqueous two-phase systems involve thermoreversible reaction–separation processes in the nucleophilic degradation of diazinon and further separation of the reaction products.
Daniela Millan; Mafalda R. Almeida; Ana F. C. S. Rufino; João A. P. Coutinho; Mara G. Freire. Nucleophilic degradation of diazinon in thermoreversible polymer–polymer aqueous biphasic systems. Physical Chemistry Chemical Physics 2021, 23, 4133 -4140.
AMA StyleDaniela Millan, Mafalda R. Almeida, Ana F. C. S. Rufino, João A. P. Coutinho, Mara G. Freire. Nucleophilic degradation of diazinon in thermoreversible polymer–polymer aqueous biphasic systems. Physical Chemistry Chemical Physics. 2021; 23 (7):4133-4140.
Chicago/Turabian StyleDaniela Millan; Mafalda R. Almeida; Ana F. C. S. Rufino; João A. P. Coutinho; Mara G. Freire. 2021. "Nucleophilic degradation of diazinon in thermoreversible polymer–polymer aqueous biphasic systems." Physical Chemistry Chemical Physics 23, no. 7: 4133-4140.
Artemisinin is a sesquiterpenoid lactone peroxide, known for its potent antimalarial activity that can be extracted from Artemisia annua L. This compound is only sparingly soluble in water, making its extraction using environmental-friendly and non-toxic aqueous solvents difficult. In the attempt to overcome this limitation, hydrotropes, which are a class of compounds that can assist in increasing the solubility of hydrophobic solutes in water, were investigated in this work. In particular, the hydrotropic capability of ionic liquids (ILs) on the aqueous solubility of artemisinin was studied. The effects of IL concentration and anion nature of 1-butyl-3-methylimidazolium-based ILs on the solubility of artemisinin at 303.2 K in water were evaluated. It is here shown the excellent capacity of ILs containing thiocyanate or dicyanamide anions to enhance the solubility of artemisinin in aqueous media, with a magnitude comparable to that obtained with the best organic solvents. Furthermore, solvatochromic parameters of the ILs aqueous solutions were also measured and combined with COSMO-RS and the cooperative hydrotropy model to establish relations between the artemisinin solubility enhancement and the solvent characteristics. The solubility enhancement of artemisinin is favored by the apolarity of the medium and the lower hydrogen-bond acceptor character of the hydrotrope.
Isabela Sales; Dinis O. Abranches; Pedro Costa; Tânia E. Sintra; Sónia P.M. Ventura; Silvana Mattedi; João A.P. Coutinho; Mara G. Freire; Simão P. Pinho. Enhancing Artemisinin Solubility in Aqueous Solutions: Searching for Hydrotropes based on Ionic Liquids. Fluid Phase Equilibria 2021, 534, 112961 .
AMA StyleIsabela Sales, Dinis O. Abranches, Pedro Costa, Tânia E. Sintra, Sónia P.M. Ventura, Silvana Mattedi, João A.P. Coutinho, Mara G. Freire, Simão P. Pinho. Enhancing Artemisinin Solubility in Aqueous Solutions: Searching for Hydrotropes based on Ionic Liquids. Fluid Phase Equilibria. 2021; 534 ():112961.
Chicago/Turabian StyleIsabela Sales; Dinis O. Abranches; Pedro Costa; Tânia E. Sintra; Sónia P.M. Ventura; Silvana Mattedi; João A.P. Coutinho; Mara G. Freire; Simão P. Pinho. 2021. "Enhancing Artemisinin Solubility in Aqueous Solutions: Searching for Hydrotropes based on Ionic Liquids." Fluid Phase Equilibria 534, no. : 112961.
L-asparaginase (ASNase, EC 3.5.1.1) is an aminohydrolase enzyme with important uses in the therapeutic/pharmaceutical and food industries. Its main applications are as an anticancer drug, mostly for acute lymphoblastic leukaemia (ALL) treatment, and in acrylamide reduction when starch-rich foods are cooked at temperatures above 100 °C. Its use as a biosensor for asparagine in both industries has also been reported. However, there are certain challenges associated with ASNase applications. Depending on the ASNase source, the major challenges of its pharmaceutical application are the hypersensitivity reactions that it causes in ALL patients and its short half-life and fast plasma clearance in the blood system by native proteases. In addition, ASNase is generally unstable and it is a thermolabile enzyme, which also hinders its application in the food sector. These drawbacks have been overcome by the ASNase confinement in different (nano)materials through distinct techniques, such as physical adsorption, covalent attachment and entrapment. Overall, this review describes the most recent strategies reported for ASNase confinement in numerous (nano)materials, highlighting its improved properties, especially specificity, half-life enhancement and thermal and operational stability improvement, allowing its reuse, increased proteolysis resistance and immunogenicity elimination. The most recent applications of confined ASNase in nanomaterials are reviewed for the first time, simultaneously providing prospects in the described fields of application.
João C. F. Nunes; Raquel O. Cristóvão; Mara G. Freire; Valéria C. Santos-Ebinuma; Joaquim L. Faria; Cláudia G. Silva; Ana P. M. Tavares. Recent Strategies and Applications for l-Asparaginase Confinement. Molecules 2020, 25, 5827 .
AMA StyleJoão C. F. Nunes, Raquel O. Cristóvão, Mara G. Freire, Valéria C. Santos-Ebinuma, Joaquim L. Faria, Cláudia G. Silva, Ana P. M. Tavares. Recent Strategies and Applications for l-Asparaginase Confinement. Molecules. 2020; 25 (24):5827.
Chicago/Turabian StyleJoão C. F. Nunes; Raquel O. Cristóvão; Mara G. Freire; Valéria C. Santos-Ebinuma; Joaquim L. Faria; Cláudia G. Silva; Ana P. M. Tavares. 2020. "Recent Strategies and Applications for l-Asparaginase Confinement." Molecules 25, no. 24: 5827.
Amongst the several processes investigated in the Biotechnology field, aqueous biphasic systems (ABS) have been a target of relevant research, particularly in the downstream processing of value-added compounds of biological nature and, more recently, in emerging applications such as in the development of artificial cells and synthetic biology, micropatterning and 3D printing, and sample pre-treatment and diagnosis. This review overviews and discusses the recent progress achieved in downstream processes employing ABS, namely by the development of integrated and continuous processes, and in the emerging applications where ABS seem to have strong potential, namely in cellular micropatterning and 3D printing, and in the pre-treatment of biological samples to improve diagnosis. Advantages and bottlenecks of ABS in all these applications are provided and discussed.
Flávia F. Magalhães; Ana P.M. Tavares; Mara G. Freire. Advances in aqueous biphasic systems for biotechnology applications. Current Opinion in Green and Sustainable Chemistry 2020, 27, 100417 .
AMA StyleFlávia F. Magalhães, Ana P.M. Tavares, Mara G. Freire. Advances in aqueous biphasic systems for biotechnology applications. Current Opinion in Green and Sustainable Chemistry. 2020; 27 ():100417.
Chicago/Turabian StyleFlávia F. Magalhães; Ana P.M. Tavares; Mara G. Freire. 2020. "Advances in aqueous biphasic systems for biotechnology applications." Current Opinion in Green and Sustainable Chemistry 27, no. : 100417.
Solubility, bioavailability, permeation, polymorphism, and stability concerns associated to solid-state pharmaceuticals demand for effective solutions. To overcome some of these drawbacks, ionic liquids (ILs) have been investigated as solvents, reagents, and anti-solvents in the synthesis and crystallization of active pharmaceutical ingredients (APIs), as solvents, co-solvents and emulsifiers in drug formulations, as pharmaceuticals (API-ILs) aiming liquid therapeutics, and in the development and/or improvement of drug-delivery-based systems. The present review focuses on the use of ILs in the pharmaceutical field, covering their multiple applications from pharmaceutical synthesis to drug delivery. The most relevant research conducted up to date is presented and discussed, together with a critical analysis of the most significant IL-based strategies in order to improve the performance of therapeutics and drug delivery systems.
Sónia Pedro; Carmen R. Freire; Armando Silvestre; Mara Freire. The Role of Ionic Liquids in the Pharmaceutical Field: An Overview of Relevant Applications. International Journal of Molecular Sciences 2020, 21, 8298 .
AMA StyleSónia Pedro, Carmen R. Freire, Armando Silvestre, Mara Freire. The Role of Ionic Liquids in the Pharmaceutical Field: An Overview of Relevant Applications. International Journal of Molecular Sciences. 2020; 21 (21):8298.
Chicago/Turabian StyleSónia Pedro; Carmen R. Freire; Armando Silvestre; Mara Freire. 2020. "The Role of Ionic Liquids in the Pharmaceutical Field: An Overview of Relevant Applications." International Journal of Molecular Sciences 21, no. 21: 8298.
Deoxyribonucleic acid (DNA) carries the genetic information essential for the growth and functioning of living organisms, playing a significant role in life sciences research. However, the long-term storage and preservation of DNA, while ensuring its bioactivity, are still current challenges to overcome. In this work, aqueous solutions of ionic liquids (ILs) were investigated as potential preservation media for double stranded (dsDNA). A screening of several ILs, by combining the cholinium, tetrabutylammonium, tetrabutylphosphonium, and 1-ethyl-3-methylimidazolium, cations with the anions bromide, chloride, dihydrogen phosphate, acetate, and glycolate, was carried out in order to gather fundamental knowledge on the molecular features of ILs that improve the dsDNA stability. Different IL concentrations and the pH effect were also addressed. Circular dichroism (CD) spectroscopy was used to evaluate the conformational structure and stability of dsDNA. IL-DNA interactions were appraised by UV-Vis absorption spectrophotometry and 31P nuclear magnetic resonance (NMR) spectroscopy. The results obtained demonstrate that pH has a significant effect towards the dsDNA stability. Amongst the ILs investigated, cholinium-based ILs are the most promising class of ILs to preserve the dsDNA structure, in which electrostatic interactions between the cholinium cation and the DNA phosphate groups play a significant role as demonstrated by the 31P NMR data, being more relevant at higher IL concentrations. On the other hand, the denaturation of dsDNA mainly occurs with ILs composed of more hydrophobic cations and able to establish dispersive interactions with the nucleobases environment. Furthermore, the IL anion has a weaker impact when compared to the IL cation effect to interact with DNA molecules. The experimental data of this work provide relevant fundamental knowledge for the application of ILs in the preservation of nucleic acids, being of high relevance in the biotechnology field.
Teresa B. V. Dinis; Fani Sousa; Mara G. Freire. Insights on the DNA Stability in Aqueous Solutions of Ionic Liquids. Frontiers in Bioengineering and Biotechnology 2020, 8, 547857 .
AMA StyleTeresa B. V. Dinis, Fani Sousa, Mara G. Freire. Insights on the DNA Stability in Aqueous Solutions of Ionic Liquids. Frontiers in Bioengineering and Biotechnology. 2020; 8 ():547857.
Chicago/Turabian StyleTeresa B. V. Dinis; Fani Sousa; Mara G. Freire. 2020. "Insights on the DNA Stability in Aqueous Solutions of Ionic Liquids." Frontiers in Bioengineering and Biotechnology 8, no. : 547857.
Prostate specific antigen (PSA) is the most widely used clinical biomarker for the diagnosis and monitoring of prostate cancer. Most available techniques for PSA quantification in human fluids require extensive sample processing and expensive immunoassays that are often unavailable in developing countries. The quantification of PSA in serum is the most common practice; however, PSA is also present in human urine, although less used in diagnosis. Herein we demonstrate the use of ionic-liquid-based aqueous biphasic systems (IL-based ABS) as effective pre-treatment strategies of human urine, allowing the PSA detection and quantification by more expedite equipment in a non-invasive matrix. If properly designed, IL-based ABS afford the simultaneous extraction and concentration of PSA (at least up to 250-fold) in the IL-rich phase. The best ABS not only allow to concentrate PSA but also other forms of PSA, which can be additionally quantified, paving the way to their use in differential prostate cancer diagnosis.
Matheus M. Pereira; João D. Calixto; Ana C. A. Sousa; Bruno J. Pereira; Álvaro S. Lima; João A. P. Coutinho; Mara G. Freire. Towards the differential diagnosis of prostate cancer by the pre-treatment of human urine using ionic liquids. Scientific Reports 2020, 10, 1 -8.
AMA StyleMatheus M. Pereira, João D. Calixto, Ana C. A. Sousa, Bruno J. Pereira, Álvaro S. Lima, João A. P. Coutinho, Mara G. Freire. Towards the differential diagnosis of prostate cancer by the pre-treatment of human urine using ionic liquids. Scientific Reports. 2020; 10 (1):1-8.
Chicago/Turabian StyleMatheus M. Pereira; João D. Calixto; Ana C. A. Sousa; Bruno J. Pereira; Álvaro S. Lima; João A. P. Coutinho; Mara G. Freire. 2020. "Towards the differential diagnosis of prostate cancer by the pre-treatment of human urine using ionic liquids." Scientific Reports 10, no. 1: 1-8.
Nucleic acids are relevant biopolymers in therapy and diagnosis, for which their purity and biological activity are of crucial relevance. However, these features are difficult to achieve by cost-effective methods. Herein, we report the functionalization of a macroporous chromatographic support functionalized with an ionic liquid (IL) with remarkable performance to purify nucleic acids. An initial screening with distinct IL chemical structures supported in silica was carried out, allowing to identify the IL 1-methyl-3-propylimidazolium chloride as the most promising ligand. A chromatographic macroporous matrix able to be used in preparative liquid chromatography was then functionalized and binding/elution studies were performed. The IL 1-methyl-3-propylimidazolium chloride acts as a multimodal ligand with a remarkable dynamic binding capacity. This macroporous support allows the (one-step) purification of nucleic acids, namely small RNAs, ribosomal RNA and genomic DNA, from a bacterial lysate, and can be regenerated and reused without compromising its separation performance.
M.C. Neves; P. Pereira; A.Q. Pedro; J.C. Martins; T. Trindade; J.A. Queiroz; M.G. Freire; F. Sousa. Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step. Materials Today Bio 2020, 8, 100086 .
AMA StyleM.C. Neves, P. Pereira, A.Q. Pedro, J.C. Martins, T. Trindade, J.A. Queiroz, M.G. Freire, F. Sousa. Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step. Materials Today Bio. 2020; 8 ():100086.
Chicago/Turabian StyleM.C. Neves; P. Pereira; A.Q. Pedro; J.C. Martins; T. Trindade; J.A. Queiroz; M.G. Freire; F. Sousa. 2020. "Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step." Materials Today Bio 8, no. : 100086.
L-Asparaginase immobilization by adsorption over MWCNTs for potential application in pharmaceutical and food industries.
Raquel O. Cristóvão; Mafalda R. Almeida; Maria A. Barros; João C. F. Nunes; Rui Boaventura; José M. Loureiro; Joaquim L. Faria; Márcia C. Neves; Mara G. Freire; Valéria C. Ebinuma-Santos; Ana P. M. Tavares; Cláudia G. Silva. Development and characterization of a novel l-asparaginase/MWCNT nanobioconjugate. RSC Advances 2020, 10, 31205 -31213.
AMA StyleRaquel O. Cristóvão, Mafalda R. Almeida, Maria A. Barros, João C. F. Nunes, Rui Boaventura, José M. Loureiro, Joaquim L. Faria, Márcia C. Neves, Mara G. Freire, Valéria C. Ebinuma-Santos, Ana P. M. Tavares, Cláudia G. Silva. Development and characterization of a novel l-asparaginase/MWCNT nanobioconjugate. RSC Advances. 2020; 10 (52):31205-31213.
Chicago/Turabian StyleRaquel O. Cristóvão; Mafalda R. Almeida; Maria A. Barros; João C. F. Nunes; Rui Boaventura; José M. Loureiro; Joaquim L. Faria; Márcia C. Neves; Mara G. Freire; Valéria C. Ebinuma-Santos; Ana P. M. Tavares; Cláudia G. Silva. 2020. "Development and characterization of a novel l-asparaginase/MWCNT nanobioconjugate." RSC Advances 10, no. 52: 31205-31213.
Since the development of liquid-phase microextraction (LPME), different LPME modes depending on the experimental set-up to carry out the extraction have been described. Dispersive liquid-liquid microextraction (DLLME), in which a small amount of the water-insoluble extraction solvent is dispersed in the sample, is the most successful mode in terms of number of applications reported. Advances within DLLME have been mainly shifted to the incorporation of green, smart and tunable materials as extraction solvents to improve the sustainability and efficiency of the method. In this sense, hydrophilic media represent a promising alternative since the water-miscibility of these substances increases the mass transfer of the analytes to the extraction media, leading to higher extraction efficiencies. Considering the variety of hydrophilic media that have been incorporated in LPME approaches resembling DLLME, this review aims to classify these methods in order to clarify the confusing terminology used for some of the strategies. Hydrophilic media covered in this review comprise surfactants, polar organic solvents, deep eutectic solvents, ionic liquids, water-miscible polymers, and switchable solvents. Different physicochemical mechanisms of phase separation are discussed for each LPME method, including the coacervation phenomena and other driving forces, such as pH, temperature, salting-out effect, metathesis reaction and organic solvents. LPME modes are classified (in cloud-point extraction, coacervative extraction, aqueous biphasic systems, and different DLLME modes depending on the extraction medium) according to both the nature of the water-miscible extraction phase and the driving force of the separation. In addition, the main advances and analytical applications of these methods in the last three years are described.
Idaira Pacheco-Fernández; Raúl González-Martín; Francisca Silva; Mara G. Freire; Verónica Pino. Insights into coacervative and dispersive liquid-phase microextraction strategies with hydrophilic media – A review. Analytica Chimica Acta 2020, 1143, 225 -249.
AMA StyleIdaira Pacheco-Fernández, Raúl González-Martín, Francisca Silva, Mara G. Freire, Verónica Pino. Insights into coacervative and dispersive liquid-phase microextraction strategies with hydrophilic media – A review. Analytica Chimica Acta. 2020; 1143 ():225-249.
Chicago/Turabian StyleIdaira Pacheco-Fernández; Raúl González-Martín; Francisca Silva; Mara G. Freire; Verónica Pino. 2020. "Insights into coacervative and dispersive liquid-phase microextraction strategies with hydrophilic media – A review." Analytica Chimica Acta 1143, no. : 225-249.