Synthesis of an antibody-like material for the detection of Albumin

6th Graduate Student Symposium on Molecular Imprinting6th Graduate Student Symposium on Molecular Imprinting, Medway School of Pharmacy, Kent, 27-28 de Agosto 2015A novel molecularly imprinted polymer (MIP) is presented for the detection of Albumin, currently a biomarker of several diseases. The material acted as an antibody for Albumin and was obtained through a bulk imprinting approach, by electropolymerizing Eriochrome blackT (EBT) around the target protein

A novel printed 3-electrode system for the electrochemical detection of sulfadiazine

We thank the financial support of 3C´s – Cellulose and Cork in the Control of antibiotics in aquaculture (PTDC/AAG-TEC/5400/2014 and POCI-01-0145-FEDER-016637), to FEDER, through COMPETE2020, POCI, and FCT (Fundação para a Ciência e a Tecnologia I.P.).info:eu-repo/semantics/publishedVersio

Purification, characterization and clinical applications of therapeutic fungal enzymes

This book chapter presents an overview of therapeutic fungal enzymes and their developments in biopharmaceuticals for the treatment of several diseases, clinical applications and investigation. Enzymes are biocatalysts of many reactions with widespread use in the pharmaceutical industry and medicine. Due to their high specificity, greater affinity, and high catalytic efficiency, enzymes have been widely used for therapeutic purposes. More specifically, therapeutic enzymes are being used in the treatment of several diseases, such as leukemia, cancer, pancreatic disorders, etc. For instance, L-asparaginase, which presents antineoplastic properties, has been used for the treatment of leukemia, namely acute lymphoblastic leukemia. Nowadays, more than 50% of the enzymes are produced by fungal sources, including the therapeutic enzymes, due to the advantages of being an economically feasible and consistent process, since it has high yield and is easy for modification and optimization of new therapeutic products. In this book chapter, readers from academies, research institutes and industries will gain useful information and in-deep knowledge on the emerging therapeutic fungal enzymes, their purification processes, characterization and medical applications.publishe

New liquid supports in the development of integrated platforms for the reuse of oxidative enzymes and polydopamine production

Polydopamine (PDA), a bioinspired polymer from mussel adhesive proteins, has attracted impressive attention as a novel coating for (nano) materials with an adequate conformal layer and adjustable thickness. Currently, PDA is obtained from dopamine chemical oxidation under alkaline conditions, limiting its use in materials sensible to alkaline environments. Envisaging a widespread use of PDA, the polymerization of dopamine by enzymatic catalysis allows the dopamine polymerization in a large range of pHs, overcoming thus the limitations of conventional chemical oxidation. Moreover, the conventional method of polymerization is a time-consuming process and produces PDA films with poor stability, which restricts its applications. On the other hand, the main bottleneck of enzyme-based biocatalytic processes is the high cost of the single use of the enzyme. In this work, laccase was used to catalyse dopamine polymerization. To improve its performance, a liquid support for integrating the laccase and its reuse together with the PDA production and recovery was developed using aqueous biphasic systems (ABS). Firstly, dopamine polymerization by laccase was optimized in terms of pH, temperature and initial dopamine concentration. It was demonstrated that the highest enzymatic polymerization of dopamine was achieved at pH 5.5, 30°C and 2 mg ml−1 of dopamine. Then, ABS composed of polymers, salts and ionic liquids were evaluated to optimize the laccase confinement in one phase while PDA is recovered in the opposite phase. The most promising ABS allowing the separation of laccase from the reaction product is composed of polypropylene glycol (400 g mol−1) and K2HPO4. The polymerization of dopamine in ABS leads to a remarkable improvement of polymerization of 3.9-fold in comparison to the conventional chemical PDA polymerization. The phase containing the confined laccase was reused for four consecutive reaction cycles, with a relative polymerization of 68.9% in the last cycle. The results of this work proved that ABS are a promising approach to create a liquid support for enzyme reuse allowing the process intensification efforts. The use of biocatalysts in ABS emerges as sustainable and alternative platforms from environmental and techno-economic points of view.publishe

Efficient extraction of the RuBisCO enzyme from spinach leaves using aqueous solutions of biocompatible ionic liquids

Ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO) is the most abundant protein on the planet, being present in plants, algae and various species of bacteria, with application in the pharmaceutical, chemical, cosmetic and food industries. However, current extraction methods of RuBisCO do not allow high yields of extraction. Therefore, the development of an efficient and selective RuBisCOs’ extraction method is required. In this work, aqueous solutions of biocompatible ionic liquids (ILs), i.e., ILs derived from choline and analogues of glycine-betaine, were applied in the RuBisCO’s extraction from spinach leaves. Three commercial imidazolium-based ILs were also investigated for comparison purposes. To optimize RuBisCO’s extraction conditions, response surface methodology was applied. Under optimum extraction conditions, extraction yields of 10.92 and 10.57 mg of RuBisCO/g of biomass were obtained with the ILs cholinium acetate ([Ch][Ac]) and cholinium chloride ([Ch]Cl), respectively. Circular dichroism (CD) spectroscopy results show that the secondary structure of RuBisCO is better preserved in the IL solutions when compared to the commonly used extraction solvent. The obtained results indicate that cholinium-based ILs are a promising and viable alternative for the extraction of RuBisCO from vegetable biomass.publishe

An Overview on the Recent Advances in Alternative Solvents as Stabilizers of Proteins and Enzymes

Currently, the use of alternative solvents is increasing, namely ionic liquids (ILs) and deep eutectic solvents (DESs) in diverse fields of knowledge, such as biochemistry, chemistry, chemical engineering, biotechnology and biomedicine. Particularly, when compared to traditional solvents, these alternative solvents have great importance for biomolecules due to the enhanced solubility, structure stability and the biological activity of biomolecules, such as protein and enzymes. Thus, in this review article, the recent developments and efforts on the technological developments carried out with ILs and DESs for the stabilization and activation of proteins and enzymes are provided. The most studied IL-and DES-based formulations for proteins and enzymes are discussed and the molecular mechanisms and interactions related to the increased stability promoted by these alternative solvents are disclosed, while emphasizing their main advantagespublishe

Directional outlyingness applied to distances between genomic words

The detection of outlier curves/images is crucial in many areas, such as environmental, meteorological, medical, or economic contexts. In the functional framework, outlying observations are not only those that contain atypically high or low values, but also curves that present a different shape or pattern from the rest of the curves in the sample. In this short paper, we mention some recent methods for outlier detection in functional data and apply a recently proposed measure, the directional outlyingness, and the functional outlier map to detect words with outlying distance distribution in the human genome.publishe

THE IMPACT OF INFORMAL CAREGIVERS DURING CANCER PATIENTS’ TRAJECTORY IN ECONOMIC HEALTH OUTCOMES

In the trajectory of the person with cancer, the presence of uncontrolled symptoms are the main and frequent cause of demand for health services, which increases with the advanced stage of the disease. The modernization of health systems advocates the domiciliation of health care, with proximity assistance, where informal caregivers assume a role of relevance. Persons with cancer prefer this typology of service, as well have positive impact in economic health outcomes. However, it is essential to reflect in the shapes of the professional support, health policies and incentives, which are needed with the finality to prevent burnout of the caregiver. As ensure the resources to certify the self-care and quality of life of the person with cancer and informal caregivers

Sustainable liquids support as a media for biocatalytic reactions using aqueous biphasic systems

Over the last few years a significant interest in the application of biocatalyst in several industries as an alternative to the commonly used chemical biocatalyst has emerged, since enzymes, such as laccase, allow for an improved, sustainable and biodegradable catalytic process partially due to their high specificity to its substrates. Nevertheless, its application can still be considered a costly process thus to overcome this shortcoming emerges the need for the enzyme reutilization employed in the catalytic reaction 1. One of the alternatives is to implement aqueous biphasic systems (ABS), which are a biocompatible liquid-liquid extraction as a liquid support. Moreover, the introduction of ionic liquids (ILs) in ABS have shown promising results in the system overall performance and could potentially allow for an improved biocatalyst performance. Therefore, our goal was to form an ABS composed of cholinium-based ILs and polypropylene glycol (PPG 400) as novel liquid supports for enzymes, using laccase as models in order to evaluated the degradation of the textile dye Remazol Brilliant Blue R (RBBR) as well as the capacity to reutilized the biocatalyst for further applications.publishe

Integrated platform resorting to ionic liquids comprising the extraction, purification and preservation of DNA

The large-scale production of therapeutically targeted-deoxyribonucleic acid (DNA) has passed through several challenges, postponing the tangible implementation of an effective, economic and sustainable manufacturing system. Such challenges comprise the need to develop an integrative downstream process able to extract, purify and long-term preserve DNA, whilst reducing the risk of degradation by endonucleases that would compromise their effectiveness as therapeutic products. In this work, three-phase partitioning (TPP) systems formed by the application of aqueous biphasic systems (ABS) composed of several biocompatible cholinium-based ionic liquids (ILs), are proposed for the separation of double stranded DNA (dsDNA) from the endonuclease deoxyribonuclease I (DNase I). By taking advantage of the tailor-made properties of ILs, dsDNA can be completely extracted to the IL-rich phase, whereas DNase I is precipitated at the ABS interphase. The ABS/TPP formed by IL cholinium glycolate ([N 111(2OH) ][Gly]) fulfills the aim of this work, i.e. at ensuring the technical viability of IL-based ABS/ TPP for the “one-pot” extraction, purification and long-term preservation of dsDNA. The results reveal the potential of this system to be applied in the bioprocessing of DNA, particularly relevant when envisioning DNA- based therapeutic products.publishe