Prototipo para la aplicación de campos eléctricos integrado a un sistema de cámaras microfluídicas para la lisis celular de bacterias productoras de PHA

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dc.contributor.advisorMerchán Castellanos, Nuri Andrea
dc.contributor.advisorOliveros Hincapié, Jorge Armando
dc.contributor.authorFrasica Romero, Karen Valentina
dc.date.accessioned2024-11-30T02:08:49Z
dc.date.available2024-11-30T02:08:49Z
dc.date.issued2024-11
dc.description.abstractLos polihidroxialcanoatos (PHAs), son un tipo de poliéster alifático sintetizado naturalmente por algunas bacterias y que presenta un gran potencial biotecnológico como bioplástico debido a que tiene propiedades fisicoquímicas similares a los plásticos de origen petroquímico. Si bien se tiene un gran interés en su producción, el costo en el proceso de extracción del biopolímero utilizando solventes halogenados aumenta su precio final en un 50%. La extracción eléctrica se presenta como una alternativa a la extracción con solventes. Es así, como Rojas diseñó e implementó un prototipo para la extracción de PHA por campos eléctricos. Pese a lograr obtener el biopolímero, la falta de control de flujo durante el procesamiento de las bacterias en el dispositivo ocasiona adherencia del material biológico a los electrodos, así como contaminación por parte de restos de oxidación de los electrodos de cobre. El objetivo de este proyecto fue el desarrollo de un prototipo para la aplicación de campos eléctricos integrado a un sistema de cámaras microfluídicas para la lisis celular de bacterias productoras de PHA. Se definió la geometría de las cámaras microfluídicas a través de una revisión bibliográfica, y mediante matrices de decisión se seleccionó la geometría rectangular. Utilizando el software COMSOL Multiphysics se simuló el campo eléctrico, permitiendo caracterizar el campo eléctrico generado por diferentes voltajes aplicados, desde los 180 V hasta los 390 V, generando un campo eléctrico de entre 150 a 1.2 Kv/m. Se realizó la simulación del comportamiento de velocidad y presión del fluido dentro de las cámaras microfluídicas, evidenciando que la geometría transversal rectangular presentó un campo eléctrico uniforme y en conjunto de la bifurcación en Y, aseguró un flujo laminar. En la fase de construcción, por técnica de transferencia termodinámica de Tóner y grabado de PCB, se obtuvieron electrodos en cobre de 15 µ𝑚 de espesor y microcámaras en PDMS que pueden procesar 3,98 µ𝐿 por canal con una velocidad aproximada de 0,06 m/s. Estas estructuras finales presentaron un error relativo de 22,47% con respecto a las medidas establecidas en el diseño detallado. Finalmente, se estableció el voltaje de operación para la extracción de Cupriavidus necator en 180 V y para Bacillus licheniformis en 285 V, de acuerdo a la relación que se identificó entre sus características morfológicas y el potencial de membrana inducido por acción del campo eléctrico. El porcentaje de extracción alcanzado con el dispositivo para Cupriavidus necator fue de 62,9% y para Bacillus licheniformis de 75,6% con un procesamiento de 2 mL en 45 segundos. Los porcentajes se compararon con los resultados obtenidos mediante el método tradicional por extracción de dispersión de hipoclorito de sodio-cloroformo. Se concluyó que la implementación de cámaras microfluídicas al prototipo de campos eléctricos, presentó ventajas tanto en la concentración del campo eléctrico como en el control de flujo, control de tiempo y aumento del volumen de procesamiento. Este trabajo fue desarrollado en colaboración con la Universidad Federal de Minas Gerais, Brasil
dc.description.abstractenglishPolyhydroxyalkanoates (PHAs) are a type of aliphatic polyester naturally synthesized by certain bacteria and hold great biotechnological potential as bioplastics due to their physicochemical properties, which are similar to petrochemical-based plastics (Müller-Santos et al., 2021). While there is significant interest in their production, the cost of extracting the biopolymer using halogenated solvents increases its final price by 50% (Garcia-Garcia et al., 2022). Electrical extraction emerges as an alternative to solvent extraction. As such, Rojas (2022) designed and implemented a prototype for PHA extraction using electric fields. Although the biopolymer was successfully obtained, the lack of flow control during bacterial processing in the device caused biological material to adhere to the electrodes, as well as contamination from copper electrode oxidation residues. The aim of this project was to develop a prototype for applying electric fields integrated with a microfluidic chamber system for cell lysis of PHA-producing bacteria. The geometry of the microfluidic chambers was defined through a literature review, and using decision matrices, the rectangular geometry was selected. COMSOL Multiphysics software was used to simulate the electric field, allowing for characterization of the field generated by different applied voltages, ranging from 180V to 390V, producing an electric field of between 150 and 1.2 KV/m. The fluid velocity and pressure behavior inside the microfluidic chambers was also simulated, showing that the rectangular cross-sectional geometry provides a uniform electric field and, together with the Y-bifurcation, ensures laminar flow. In the construction phase, by thermodynamic toner transfer technique and PCB etching, 15 m thick copper electrodes and PDMS microchambers were obtained that can process 3.98 L per channel with an approximate speed of 0.06 m/s. These final structures presented a relative error of 22.47% with respect to the measurements established in the detailed design. Finally, the operating voltage for the extraction of Cupriavidus necator was set at 180 V and for Bacillus licheniformis at 285 V, according to the relationship identified between their morphological characteristics and the membrane potential induced by the action of the electric field. The extraction percentage achieved with the device for Cupriavidus necator was 62.9% and for Bacillus licheniformis 75.6% with a processing of 2 mL in 45 seconds. The percentages were compared with the results obtained using the traditional method by sodium hypochlorite-chloroform dispersion extraction. It was concluded that the implementation of microfluidic chambers to the electric field prototype presented advantages both in electric field concentration and in flow control, time control and increased processing volume. This work was developed in collaboration with the Federal University of Minas Gerais, Brazil
dc.description.degreelevelPregradospa
dc.description.degreenameBioingenierospa
dc.description.sponsorshipUniversidad Federal de Minas Gerais
dc.format.mimetypeapplication/pdf
dc.identifier.instnameinstname:Universidad El Bosquespa
dc.identifier.reponamereponame:Repositorio Institucional Universidad El Bosquespa
dc.identifier.repourlhttps://repositorio.unbosque.edu.co
dc.identifier.urihttps://hdl.handle.net/20.500.12495/13491
dc.language.isoes
dc.publisher.facultyFacultad de Ingenieríaspa
dc.publisher.grantorUniversidad El Bosquespa
dc.publisher.programBioingenieríaspa
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dc.rightsAtribución-NoComercial-CompartirIgual 4.0 Internacionalen
dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.rights.accessrightshttp:/purl.org/coar/access_right/c_abf2/
dc.rights.localAcceso abiertospa
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/
dc.subjectMicrofluídica
dc.subjectPolihidroxialcanoatos
dc.subjectElectroporación
dc.subjectCampo eléctrico
dc.subject.ddc610.28
dc.subject.keywordsMicrofluidics
dc.subject.keywordsPolyhydroxyalkanoates
dc.subject.keywordsElectroporation
dc.subject.keywordsElectric field
dc.titlePrototipo para la aplicación de campos eléctricos integrado a un sistema de cámaras microfluídicas para la lisis celular de bacterias productoras de PHA
dc.title.translatedPrototype for the application of electric fields integrated to a microfluidic chamber system for cell lysis of PHA-producing bacteria
dc.type.coarhttps://purl.org/coar/resource_type/c_7a1f
dc.type.coarversionhttps://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.driverinfo:eu-repo/semantics/bachelorThesis
dc.type.hasversioninfo:eu-repo/semantics/acceptedVersion
dc.type.localTesis/Trabajo de grado - Monografía - Pregradospa

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