Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Simulation of protein adsorption in Ion Exchange Chromatography (IEX) with COMSOL Multiphysics(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-12-07) Cardoso Ortegón, Alan; IBARRA HERRERA, CELESTE CONCEPCION; 265151; Mata Gómez, Marco Arnulfo; puemcuervo; Sánchez, Mirna Lorena; Ibarra Herrera, Celeste Concepción; Cervantes Avilés, Pabel Antonio; Escuela de Ingeniería y Ciencias; Campus Puebla; Gómez Sánchez, Carlos EduardoA simulation of Ion Exchange Chromatography in COMSOL Multyphisics by the mass transport phenomena on a column with packed bed heigh of 56.52 cm. The simulation was based on the physical phenomena of convections and diffusion, both radial and axial, the chemical reaction simulated was that of Bovine serum Albumin (BSA) and recombinant Green Fluorescent Protein (rGFP) with Q Sepharose XLTM in slightly basic conditions pH 8.5. Experimental data of Langmuir Adsorption Isotherm in batch was used to determine an equilibrium constant kL and use it for the simulation reaction. Dynamic adsorption experimental data was obtained by frontal analysis of BSA and rGFP in an Äkta Pure system, that resulted in a chromatogram. The chromatogram was studied with Carta’s model to obtain physical parameters of the model proteins. The simulations were based on the experimental data and fed in several nodules of the Software to solve for a chromatographic process of adsorption through time and radial diffusion. Nodules of reaction rate of a chemical reaction (chemistry), mass transport through a system based on convection and diffusion (Transport of Dilutes Species) and for velocity vector (Darcy’s Law) were used. Coupling these nodules, the study of mass transport in a column can be visualized in 3D and the results can be plotted in a two dimensional form, this allows to plot the process in the radial direction and not only in the time dimension.
- Fabrication and characterization of an anion exchange-based monolithic column, using γ-aminobutyric acid as a ligand, contained in a 3D printed casing for separation of biomolecules(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-09-22) Ceballos Medina, Javier; Mata Gómez, Marco Arnulfo; dnbsrp; Ornelas Soto, Nancy Edith; Gordillo Guerra, Paola Guadalupe; Escuela de Ingeniería y Ciencias; Campus Monterrey; Ibarra Herrera, Celeste ConcepciónA poly(GMA-co-EDMA) monolith was synthesized within a 3D printed housing of 4 different materials and modified with γ-aminobutyric acid, a non-protein amino acid. The monoliths were synthesized with 3 different surface pretreatments in housing with 5 mm i.d. and 20 mm length channel. The functionalization of the monolith was done by by Shiff-based method using diethylenediamine as a spacer. SEM images evidenced that a homogeneous surface monolith on γ-MAPS treated surfaces while direct synthesis had scales and big agglomerates within the polymeric matrix. Raman spectroscopy evidenced the different modifications the resin housing surfaces experienced and confirmed the presence of the ligand. On the other hand, BET analysis found surface areas ranging 7.459 m2/g to 20.147 m2/g and micropores range from 0.432 to 1.736 nm, being 4.70 nm the most common pore. The fabricated monolithic columns with EDMA-G, γ-MAPS at 20%, γ-MAPS at 30% treatments exhibited ionic capacities of 0.056±0.001, 0.174±0.001 and 0.293±0.015 mmol Cl⁻/mL, respectively. EDMA-G column exhibited the best performance for fractionating proteins from a concentrated bacterial lysate from E. coli strain BL21 star with plasmid pSB21C3, producing RFP. Eventhough the column was able to retain 20% of the total RFP, improvement needs to be done to increase the ligand density and, in consequence, the adsorption capacity.
- Fabrication of a guanidine ligand-based anion exchange monolithic stationary phase in a 3D printed polypropylene housing for protein chromatography(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-01-01) Pérez Rodríguez, Elizabeth; MATA GOMEZ, MARCO ARNULFO; 207149; Mata Gómez, Marco Arnulfo; puelquio/mscuervo; Luna Vital, Diego Armando; Sánchez, Mirna Lorena; González Valdez, José Guillermo; School of Engineering and Sciences; Campus Monterrey; Ibarra Herrera, Celeste ConcepciónA monolithic anion exchange support is synthesized based on the incorporation of γ -guanidinobutyric acid an alkaloid with three resonant amino groups- as ligand onto a poly(EDMA-co-GMA) monolith. The created monolithic anion exchange support is referred as M-Gnd. Monolith was synthesized in a one-step polymerization reaction in a designed and printed polypropylene housing within a 5x20mm i.d. channel. Functionalization of the poly-methacrylate monolith with γ -guanidinobutyric acid was done by Shift-based method using diethylenediamine as a spacer. Homogeneous surface morphology was appreciated by SEM image, and FTIR analysis confirmed the presence of ligand. Highest value of Dynamic binding capacity (12.7mg BSA/mL monolith) was obtained at 0.5 mL/min when a 2 mg/mL protein concentration was used. The estimated ligand for M-Gnd was 1.23 mmol/g dry support. The M-Gnd support allowed to purity a red fluorescent protein up to 75%, presenting a high efficiency (N and HETP values). The characterized M-Gnd showed positive results as anion exchange chromatography support for protein purification. However, the functionalization of γ-guanidinobutyric acid can be improved to have a comparable chromatographic performance to commercial anion exchange monolith.
- Design of droplet-based microfluidic devices and its application for protein fractionation(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-04) Hernández Cid, David; Mata Gómez, Marco Arnulfo; qro /|bqrotbecerra/tolmquevedo; Pérez González, Victor Hugo; Cervantes Avilés, Pabel Antonio; School of Engineering and Science; Campus Monterrey; González Valdéz, José GuillermoMicrofluidics technology offers new possibilities due to the nature of small scale, such as the high surface area to volume ratio. Droplet based microfluidics is an emerging field that has found its application in different areas, like material science, chemical reactions, and biochemical analysis. On the other hand, downstream processing operations like separation and purification still has some drawbacks like big reagents consumption, huge waste generation, extra steps like concentration or salts removal. Here is where microfluidics can be implemented to overcome some of the issues presented in this field. In this work, we designed and developed a microfluidic platform able to generate droplet-like aqueous two- phases system (ATPS) in a continuous manner. We started this project by performing a study of how different forces like surface tension, inertial forces, viscous forces, and geometry affect droplet formation. We found that all the previously mentioned parameters have an impact in different ways on the process of droplet formation. Afterwards, this knowledge was used to implement a microfluidic flow-focusing on the generation of droplet-based micro ATPS with the used of salts and PEG as the two liquid phases of the ATPS. To see that this ATPS droplet microfluidic system really worked, we tested its capacity to fractionate ribonuclease A (RNase A) on its native and PEGylated forms, which have affinity for the salt and PEG phases, respectively. Thus, we expected that proteins would get inside or outside droplets or streamlines according to their affinity for each of the two phases. In doing so, native, and PEGylated proteins were labeled with FITC and their movement from one to another phase or their retention in the phase where protein was placed were recorded. It was confirmed that proteins move or keep in their phase for which they have more affinity. This result opens the possibility for the development of microfluidic separators based on ATPS droplet formation.
- Mathematical modeling of the enzymatic saccharification process of lignocellulosic biowaste(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-11) Fedeli, Vittorio; Mata Gómez, Marco Arnulfo; tolmquevedo; Calderón Oliver, Mariel; Aguilar Jiménez, Oscar Alejandro; School of Engineering and Science; Campus Monterrey; Gómez Sánchez, Carlos EduardoLignocellulose is a biowaste produced in large quantities by industries; approximately 181.5 billion tons are produced annually in the world. This makes this type of residue a qualifiable candidate resource of energy, which nowadays, is underutilized. It is estimated that the food processing industry produces around 1.3 billion tons per year. In Mexico, the craft beer industry produces 3.8 thousand tons per year of brewers' spent grain. Being Mexico's fastest-growing industry, it can be considered a suitable source of biowaste. Brewers spent grain is considered a lignocellulosic material, which possesses a complicated structure containing lignin, hemicellulose, and cellulose. Due to its complexity, diversity, and recalcitrance to degradation, specific pretreatments to degrade it have been developed, such as biological, chemical, physical, and physicochemical. Notwithstanding, in nature, fungi are well-known microorganisms capable of degrading it through a tremendous battery of enzymes that are secreted in an ordered and systematic fashion. Nonetheless, the full understanding of this process, and the order in which each enzyme acts on lignocellulose, is far to be elucidated. Therefore, the present thesis aims to develop fungi bio-inspired mechanistic mathematical model capable of describing the enzymatic degradation process of lignocellulose biomass (brewers spent grain) and evaluate it through different experiments. Sequential addition of enzymes to biowaste, as well as experiments involving the addition of a pool without key enzymes that were further added at a specific time, were evaluated. Overall, results revealed that the lignin is not the most resilient and dense layer of lignocellulose as it has been believed. On the contrary, it seems lignin forms pore-like structures and diffuses through all different layers of this substrate. When hemicellulases (xylanases and pectinases) were not present in the enzyme pool, the reaction was not favored, indicating the importance of this polymer in lignin structure. These results gave an idea of how fungi work in nature and how the polymer layers are organized in lignin. However, to fully confirm these findings, more tests need to be performed to generate a robust and proven mechanistic mathematical model, enabling us to lay the foundations of a potential industrial-scale process.
- Metabolic flux analysis of Xanthophyllomyces dendrorhous metabolism to understand the production of astaxanthin(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06) Martínez Castro, Victor Ignacio; MATA GOMEZ, MARCO ARNULFO; 207149; Mata Gómez, Marco Arnulfo; lagdtorre, emipsanchez; Benavides Lozano, Jorge Alejandro; Licona Cassani, Cuauhtémoc; School of Engineering and Sciences; Campus Toluca; Goméz Sánchez, Carlos EduardoCarotenoid production by microorganisms, contrary to chemical synthesis, could fulfill the in-creasing demand for human consumption. The yeastX. drendrorhousis one of the most promis-ing and economically attractive microorganisms for industrial production of astaxanthin. Themetabolic pathway through which this yeast synthesize this valuable carotenoid is known. How-ever, the complex mechanisms that are involved in the process, the distribution of the metabolicfluxes and the rates at which the pathways work, remain unknown. Several studies have provideddifferent approaches to manipulate and improve carotenoid production inX. dendrorhousfromclassical mutagenesis to genetic engineering of the complete pathway covering improved precur-sor supply for carotenogenesis, enhanced metabolite flow into the pathway, and manipulating therelation C/N in the culture medium. However, it has not been reported quantitatively how nutri-ents, from the central metabolism and other pathways, converge in the carotenoids biosynthesis.In this study, the metabolism ofX. dendrorhousgrowing in a continuous culture, under two am-monia conditions, Limited (L) and Non-limited (NL). The metabolic flux analysis (MFA) allowedto understand the distribution the intracellular fluxes along the different metabolic pathways eval-uated, but most important, it elucidated that by limiting the ammonia assimilation flux(L= 0.002±1.1 E-05; NL= 0.004±4.3 E-05; g/gcellh), a promotion of the astaxanthin formation flux wasobtained (L= 86.4±0.6; NL= 0; ug/gcellh). This first approach will help to set a deeper studyin order to understand the metabolic pathways that regulate the flux towards the astanxanthinbiosynthesis