Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551014
Pertenecen a esta colección Tesis y Trabajos de grado de los Doctorados correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Development of novel polymer-protein conjugates and characterization of chromatographic supports(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-06-10) Sánchez Trasviña, Calef; SANCHEZ TRASVIÑA, CALEF; 655893; Rito Palomares, Marco Antonio; emipsanchez; Aguilar Jiménez, Oscar Alejandro; Zavala Arcos, Judith; Chuck Hernández, Cristina Elizabeth; School of Engineering and Sciences; Campus Monterrey; Mayolo Deloisa, Karla PatriciaProtein versatility has positioned them as a high-value biotechnological product. Among protein applications, its use as a therapeutic agent is highlighted. However, some therapeutic proteins need a modification process to increase their pharmacokinetic properties. During the protein modification process, the purification step, mainly performed by chromatography, represents a critical stage in ensuring the safety of modified therapeutics proteins. Being chromatography the main purification method, it is mandatory to fully understand the effect of all its operational variables on the separation performance. This work presents a deep analysis of chromatographic strategies used to purify modified therapeutic proteins commercially available. Furthermore, an alternative protein modification process is presented using N-(2-hydroxypropyl) methacrylamide (HPMA) polymer and its purification by chromatography. Besides, the development and characterization of PEGylated monoliths as an alternative to purify PEGylated proteins is performed. Finally, the characterization of core-shell particles being used as chromatographic support is developed. The results showed that the selection of purification strategies of commercial modified proteins depends on physicochemical properties of both protein and attached molecule and is highly dependent on the matrix where the protein is recovered. HPMA copolymers can be conjugated with Ribonuclease A (RNase A) under non-demanding conditions (PO4-3 buffer 50 mM pH 5.1 + 20 mM NaBH3CN). The new conjugates showed higher hydrophobic behavior than the native protein being this feature exploited by hydrophobic interaction chromatography (using 1.5 M (NH4)2SO4) to separate the conjugates from the unreacted protein. On the other hand, PEGylated monoliths can separate PEGylated RNase A and even isoforms when large polyethylene glycol molecules (>20 kDa) are attached to the monolith. Lastly, the CaptoTM Core 700 resin structural (50.4 nm pore size, 4.18 µm shell thickness, and 90.7 µm particle size) and adsorptive properties allowed modeling and adsorption prediction of two model proteins. In combination, all these results represent new knowledge in the polymer-protein technology and chromatography areas that proportionate guidelines to purify a wide range of molecules such as native, recombinant, and modified proteins efficiently.
- The enhancement of the health potential of prickly pear fruits with the use of high hydrostatic pressure(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06-05) Gómez Maqueo Cerecer, Andrea; GOMEZ MAQUEO CERECER, ANDREA; 692751; Welti Chanes, Jorge; emipsanchez; Jacobo Velázquez, Daniel A.; García Cayuela, Tomás; Martín Cabrejas, María Ángeles; Hector Campanella, Osvaldo; Fornari Reale, Tiziana; Escuela de Ingeniería y Ciencias; Campus Monterrey; Cano Dolado, M. PilarPrickly pear (Opuntia ficus-indica L. Mill.) fruits are low-cost, sustainable sources of bioactive compounds which could contribute to the reduction of risk factors related to obesity and metabolic syndrome. However, for prickly pears to exert mentioned health benefits, bioactive compounds must be first released from the food matrix, transformed in the gastrointestinal tract and absorbed by our bodies. High hydrostatic pressure (HHP) is an innovative food processing technology which affects the microstructure of the foods and modifies/ruptures intracellular compartments where bioactive compounds are located. Thus, contributing to their immediate release as well as to their release in the gastrointestinal tract in early, intermediate or late stages of digestion. The aim of this dissertation was to enhance the health potential of prickly pear fruits with the use of HHP by increasing the bioaccessibility of their bioactive compounds. To achieve this goal, the following topics were thoroughly assessed: (i) the characterization and quantification of bioactive compounds (betalains, phenolic compounds, carotenoids and ascorbic acid) in different Mexican and Spanish prickly pear fruits; (ii) the antioxidant, anti-inflammatory, anti-hyperglycemic and delipidating activity of their extracts and of their main isolated bioactive compounds; (iii) the effect of HHP on target bioactive compounds in prickly pear fruits, their bioactivity and microstructure; and (iv) the in vitro digestive stability and bioaccessibility of bioactive compounds in control and HHP-treated prickly pear fruits. On one hand, pulps were studied due to their importance as the edible fraction of the fruit. Meanwhile, peels were evaluated because they may be used as by-products to obtain healthy ingredients. The major finding of this dissertation was that HHP could, in fact, enhance the bioaccessibility of bioactive compounds in prickly pear fruits, thus contributing to their health potential. The studies included in this dissertation have been carried out hoping to contribute to the use of innovative technologies for the development of healthy foods so that we can live longer and healthier lives.