Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery
In recent years, pharmaceutical field has been searching for sustainable drug delivery vehicles to treat persistent medical conditions and diseases. This project seeks to perform a physicochemical analysis for OmpA's mutants (mutant 6 and mutant 12) in order to verify their viability as transpo...
- Autores:
-
Mendoza Ospina, Jhon Esteban
Martínez Chavarro, Paula Andrea
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2020
- Institución:
- Universidad de los Andes
- Repositorio:
- Séneca: repositorio Uniandes
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.uniandes.edu.co:1992/49325
- Acceso en línea:
- http://hdl.handle.net/1992/49325
- Palabra clave:
- Plásmidos
Proteínas
Escherichia coli
Ingeniería
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by-nc-nd/4.0/
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Séneca: repositorio Uniandes |
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|
dc.title.es_CO.fl_str_mv |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
title |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
spellingShingle |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery Plásmidos Proteínas Escherichia coli Ingeniería |
title_short |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
title_full |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
title_fullStr |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
title_full_unstemmed |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
title_sort |
Computational physicochemical analysis and production of OmpA's mutants with functional potential in drug delivery |
dc.creator.fl_str_mv |
Mendoza Ospina, Jhon Esteban Martínez Chavarro, Paula Andrea |
dc.contributor.advisor.none.fl_str_mv |
González Barrios, Andrés Fernando |
dc.contributor.author.none.fl_str_mv |
Mendoza Ospina, Jhon Esteban Martínez Chavarro, Paula Andrea |
dc.contributor.jury.none.fl_str_mv |
Fernández Niño, Miguel Angel |
dc.subject.armarc.es_CO.fl_str_mv |
Plásmidos Proteínas Escherichia coli |
topic |
Plásmidos Proteínas Escherichia coli Ingeniería |
dc.subject.themes.none.fl_str_mv |
Ingeniería |
description |
In recent years, pharmaceutical field has been searching for sustainable drug delivery vehicles to treat persistent medical conditions and diseases. This project seeks to perform a physicochemical analysis for OmpA's mutants (mutant 6 and mutant 12) in order to verify their viability as transport vehicles for drug delivery. For this objective, numerous computational programs were used (I-TASSER, Swiss Model, Chimera and ExPASy Bionformatics Resource Portal) to determine physicochemical, structural and translocation properties for both mutants. On the other hand, this project pursues to standardize the methodology for expression, quantification and purification of the mentioned mutants. The above with the aim of designing a replicable process that guarantees the target proteins production. The methodology for mutants'production was based in transferring synthetized genes sequences from pUC-57 plasmid to pET6xHN-N cloning vector through digestion, ligation and transformation protocols in Escherichia coli strains. Additionally, purification and quantification process for mutant 6 was developed following affinity chromatography column and SDS page techniques. This procedure was unclear in terms of a successful purification because in polyacrylamide gel only appeared a low-resolution trace of the target bands, instead of high- resolution bands that confirm an effective purification. The results showed that both mutants have physicochemical properties (cationic- charged surface, amphipathic condition, high solubility in water and thermostability) and protein structures (an anti-parallel ? sheet that composed the secondary structure) that allow to predict an effective translocation of the cell membrane. Besides, the production pathway proposed was effective, but purification and quantification processes should be modified for obtaining more accurate results |
publishDate |
2020 |
dc.date.issued.none.fl_str_mv |
2020 |
dc.date.accessioned.none.fl_str_mv |
2021-02-18T12:48:41Z |
dc.date.available.none.fl_str_mv |
2021-02-18T12:48:41Z |
dc.type.spa.fl_str_mv |
Trabajo de grado - Pregrado |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
dc.type.content.spa.fl_str_mv |
Text |
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http://purl.org/redcol/resource_type/TP |
format |
http://purl.org/coar/resource_type/c_7a1f |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/1992/49325 |
dc.identifier.pdf.none.fl_str_mv |
u833242.pdf |
dc.identifier.instname.spa.fl_str_mv |
instname:Universidad de los Andes |
dc.identifier.reponame.spa.fl_str_mv |
reponame:Repositorio Institucional Séneca |
dc.identifier.repourl.spa.fl_str_mv |
repourl:https://repositorio.uniandes.edu.co/ |
url |
http://hdl.handle.net/1992/49325 |
identifier_str_mv |
u833242.pdf instname:Universidad de los Andes reponame:Repositorio Institucional Séneca repourl:https://repositorio.uniandes.edu.co/ |
dc.language.iso.es_CO.fl_str_mv |
eng |
language |
eng |
dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ http://purl.org/coar/access_right/c_abf2 |
eu_rights_str_mv |
openAccess |
dc.format.extent.es_CO.fl_str_mv |
20 hojas |
dc.format.mimetype.es_CO.fl_str_mv |
application/pdf |
dc.publisher.es_CO.fl_str_mv |
Uniandes |
dc.publisher.program.es_CO.fl_str_mv |
Ingeniería Química |
dc.publisher.faculty.es_CO.fl_str_mv |
Facultad de Ingeniería |
dc.publisher.department.es_CO.fl_str_mv |
Departamento de Ingeniería Química y de Alimentos |
dc.source.es_CO.fl_str_mv |
instname:Universidad de los Andes reponame:Repositorio Institucional Séneca |
instname_str |
Universidad de los Andes |
institution |
Universidad de los Andes |
reponame_str |
Repositorio Institucional Séneca |
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Repositorio Institucional Séneca |
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spelling |
Al consultar y hacer uso de este recurso, está aceptando las condiciones de uso establecidas por los autores.http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2González Barrios, Andrés Fernando025dc7b7-658a-4d8a-9451-f370e8c667fd400Mendoza Ospina, Jhon Esteban293baedb-e0f8-4cd9-880d-a4773f22a4c4500Martínez Chavarro, Paula Andrea8ce63c26-1a22-47a7-acf3-0b597787c675500Fernández Niño, Miguel Angel2021-02-18T12:48:41Z2021-02-18T12:48:41Z2020http://hdl.handle.net/1992/49325u833242.pdfinstname:Universidad de los Andesreponame:Repositorio Institucional Sénecarepourl:https://repositorio.uniandes.edu.co/In recent years, pharmaceutical field has been searching for sustainable drug delivery vehicles to treat persistent medical conditions and diseases. This project seeks to perform a physicochemical analysis for OmpA's mutants (mutant 6 and mutant 12) in order to verify their viability as transport vehicles for drug delivery. For this objective, numerous computational programs were used (I-TASSER, Swiss Model, Chimera and ExPASy Bionformatics Resource Portal) to determine physicochemical, structural and translocation properties for both mutants. On the other hand, this project pursues to standardize the methodology for expression, quantification and purification of the mentioned mutants. The above with the aim of designing a replicable process that guarantees the target proteins production. The methodology for mutants'production was based in transferring synthetized genes sequences from pUC-57 plasmid to pET6xHN-N cloning vector through digestion, ligation and transformation protocols in Escherichia coli strains. Additionally, purification and quantification process for mutant 6 was developed following affinity chromatography column and SDS page techniques. This procedure was unclear in terms of a successful purification because in polyacrylamide gel only appeared a low-resolution trace of the target bands, instead of high- resolution bands that confirm an effective purification. The results showed that both mutants have physicochemical properties (cationic- charged surface, amphipathic condition, high solubility in water and thermostability) and protein structures (an anti-parallel ? sheet that composed the secondary structure) that allow to predict an effective translocation of the cell membrane. Besides, the production pathway proposed was effective, but purification and quantification processes should be modified for obtaining more accurate results"En la actualidad, el sector farmacéutico ha buscado vehículos para transporte de medicamentos con el propósito de tratar condiciones médicas persistentes. Este proyecto busca desarrollar un análisis fisicoquímico a mutantes de la proteína OmpA (mutante 6 y 12) para verificar su viabilidad como vehículos de transporte en la liberación de medicamentos. Para esto, diversos programas computacionales fueron utilizados (I-TASSER, Swiss Model, Chimera y ExPASy Bionformatics Resource Portal) para determinar propiedades fisicoquímicas, estructurales y de translocación. Por otro lado, se busca estandarizar la metodología para la expresión, cuantificación y purificación de los mutantes mencionados. Lo anterior con el propósito de diseñar un proceso replicable que garantice la producción de las proteínas seleccionadas. La metodología para la producción se basó en la trasferencia de genes sintetizados desde el plásmido pUC-57 hacia el vector de clonación pET6xHN-N a través de protocolos de digestión, ligación y transformación en la cepa Escherichia coli. Adicionalmente, procesos de purificación y cuantificación fueron desarrollados para el mutante 6, siguiendo los protocolos de cromatografía por columna de afinidad y técnicas de SDS-page. Este procedimiento fue poco claro en términos de una purificación exitosa, ya que solo se obtuvieron trazas de bandas en baja resolución en vez de bandas bien definidas que indicaban una purificación efectiva. Los resultados mostraron que ambos mutantes tienen propiedades fisicoquímicas (superficie con potencial catiónico, condición amfipática, alta solubilidad en agua y termo-estabilidad) y estructuras proteicas (hoja - anti-paralela que compone la estructura secundaria) que permiten predecir una translocación efectiva de la membrana celular. En adición, la ruta de producción propuesta para ambos mutantes fue efectiva, pero la purificación y cuantificación proteica debe ser modificada para obtener resultados más acertados."--Tomado del Formato de Documento de GradoIngeniero QuímicoPregrado20 hojasapplication/pdfengUniandesIngeniería QuímicaFacultad de IngenieríaDepartamento de Ingeniería Química y de Alimentosinstname:Universidad de los Andesreponame:Repositorio Institucional SénecaComputational physicochemical analysis and production of OmpA's mutants with functional potential in drug deliveryTrabajo de grado - Pregradoinfo:eu-repo/semantics/bachelorThesishttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_970fb48d4fbd8a85Texthttp://purl.org/redcol/resource_type/TPPlásmidosProteínasEscherichia coliIngenieríaPublicationTHUMBNAILu833242.pdf.jpgu833242.pdf.jpgIM Thumbnailimage/jpeg34019https://repositorio.uniandes.edu.co/bitstreams/a04853c8-34e9-48e3-9a8a-73e98db3ba27/downloadb177eceaf7d64f303f2d85f5b441214dMD55TEXTu833242.pdf.txtu833242.pdf.txtExtracted texttext/plain56886https://repositorio.uniandes.edu.co/bitstreams/81cbe574-b8bf-4859-a1af-70f79b4504fb/download01d82d9969006893717b69c21277a52fMD54ORIGINALu833242.pdfapplication/pdf1313051https://repositorio.uniandes.edu.co/bitstreams/c1604bdd-efd3-4d0a-8917-4f0a8243309d/downloadde2efc560316f00b4749b0ff5fe14f8dMD511992/49325oai:repositorio.uniandes.edu.co:1992/493252023-10-10 19:07:57.304http://creativecommons.org/licenses/by-nc-nd/4.0/open.accesshttps://repositorio.uniandes.edu.coRepositorio institucional Sénecaadminrepositorio@uniandes.edu.co |