Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned

[eng] In this thesis, two recombinant Pichia pastoris strains producing chimeric HPV-HIV L1P18 protein were constructed. One intracellularly produces the chimeric protein while the other secrets it. For the former, several purification methods were systematically tested and the purification process...

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Detalles Bibliográficos
Autor: Eto, Yoshiki
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/200812
Acceso en línea:https://hdl.handle.net/2445/200812
http://hdl.handle.net/10803/688692
Access Level:acceso abierto
Palabra clave:Papil·lomavirus
Vacuna del papil·lomavirus
VIH (Virus)
Llevats
Papillomaviruses
Papillomavirus vaccines
HIV (Viruses)
Yeast
id ES_c48d0cea11bcd4ccdbe5030a2d2eb5b8
oai_identifier_str oai:diposit.ub.edu:2445/200812
network_acronym_str ES
network_name_str España
repository_id_str
spelling Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learnedEto, YoshikiPapil·lomavirusVacuna del papil·lomavirusVIH (Virus)LlevatsPapillomavirusesPapillomavirus vaccinesHIV (Viruses)Yeast[eng] In this thesis, two recombinant Pichia pastoris strains producing chimeric HPV-HIV L1P18 protein were constructed. One intracellularly produces the chimeric protein while the other secrets it. For the former, several purification methods were systematically tested and the purification process was optimized. After cell lysis, virus like particles (VLPs) from L1P18 protein were purified by a combination of ammonium sulfate precipitation, size exclusion chromatography, ultracentrifugation and ultrafiltration. At the end of the purification process, the chimeric VLPs were recovered with 96% purity and 9.2% overall yield, and the morphology of VLPs were confirmed by transmission electron microscopy. This work contributes towards the development an alternative platform for production of a bivalent vaccine against HPV and HIV in P. pastoris.Universitat de BarcelonaJoseph Munné, JoanAligué i Alemany, Rosa MariaUniversitat de Barcelona. Facultat de Medicina2021info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/200812http://hdl.handle.net/10803/688692Tesis Doctorals - Facultat - Medicinareponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaIngléscc by-nc (c) Eto, Yoshiki, 2023http://creativecommons.org/licenses/by-nc/3.0/es/info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2008122026-05-27T06:46:51Z
dc.title.none.fl_str_mv Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
title Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
spellingShingle Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
Eto, Yoshiki
Papil·lomavirus
Vacuna del papil·lomavirus
VIH (Virus)
Llevats
Papillomaviruses
Papillomavirus vaccines
HIV (Viruses)
Yeast
title_short Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
title_full Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
title_fullStr Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
title_full_unstemmed Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
title_sort Production, purification and characterization of virus-like particles from chimeric HPV-HIV L1P18 protein expressed in Pichia pastoris: Lessons learned
dc.creator.none.fl_str_mv Eto, Yoshiki
author Eto, Yoshiki
author_facet Eto, Yoshiki
author_role author
dc.contributor.none.fl_str_mv Joseph Munné, Joan
Aligué i Alemany, Rosa Maria
Universitat de Barcelona. Facultat de Medicina
dc.subject.none.fl_str_mv Papil·lomavirus
Vacuna del papil·lomavirus
VIH (Virus)
Llevats
Papillomaviruses
Papillomavirus vaccines
HIV (Viruses)
Yeast
topic Papil·lomavirus
Vacuna del papil·lomavirus
VIH (Virus)
Llevats
Papillomaviruses
Papillomavirus vaccines
HIV (Viruses)
Yeast
description [eng] In this thesis, two recombinant Pichia pastoris strains producing chimeric HPV-HIV L1P18 protein were constructed. One intracellularly produces the chimeric protein while the other secrets it. For the former, several purification methods were systematically tested and the purification process was optimized. After cell lysis, virus like particles (VLPs) from L1P18 protein were purified by a combination of ammonium sulfate precipitation, size exclusion chromatography, ultracentrifugation and ultrafiltration. At the end of the purification process, the chimeric VLPs were recovered with 96% purity and 9.2% overall yield, and the morphology of VLPs were confirmed by transmission electron microscopy. This work contributes towards the development an alternative platform for production of a bivalent vaccine against HPV and HIV in P. pastoris.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
info:eu-repo/semantics/publishedVersion
format doctoralThesis
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/200812
http://hdl.handle.net/10803/688692
url https://hdl.handle.net/2445/200812
http://hdl.handle.net/10803/688692
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv cc by-nc (c) Eto, Yoshiki, 2023
http://creativecommons.org/licenses/by-nc/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc by-nc (c) Eto, Yoshiki, 2023
http://creativecommons.org/licenses/by-nc/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universitat de Barcelona
publisher.none.fl_str_mv Universitat de Barcelona
dc.source.none.fl_str_mv Tesis Doctorals - Facultat - Medicina
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869418891069882368
score 15,300724

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