Pre-clinical development of chimeric virus-like particles based HPV: HIV vaccines by using mammalian cell expression system: limitations and challenges

dc.contributor
Universitat de Barcelona. Facultat de Medicina
dc.contributor.author
Chen, Chun-Wei
dc.date.accessioned
2023-07-21T14:28:33Z
dc.date.available
2023-07-21T14:28:33Z
dc.date.issued
2022-11-08
dc.identifier.uri
http://hdl.handle.net/10803/688790
dc.description
Programa de Doctorat en Biomedicina / Tesi realitzada al Vall d'Hebron Institut de Recerca (VHIR)
ca
dc.description.abstract
[eng] HPV and HIV-1 are important public health issues in some developing and industrialized countries, but an effective chimeric HPV:HIV preventive vaccine is still unachievable. We aimed to establish an alternative mammalian (293F) cell expression system combining with chromatographic purification methods to reach an appreciable expression level, purity and recovery rate of chimeric HPV:HIV VLPs. In this study, the chimeric HPV:HIV (L1:P18I10 and L1:T20) immunogens were designed and produced by using 293F expression system. The HPV:HIV VLPs were subsequently purified by a 3-step chromatographic method, including cation (CEC), size exclusion (SEC) and heparin affinity (H-AC) chromatography. Then, the in vitro stability, in vitro self assembly and morphology of purified HPV:HIV VLPs were confirmed by non-reducing SDS-PAGE, molecular mass assay, transmission electron microscopy (TEM) respectively. The sequential and conformational P18I10 and T20 peptides presented on chimeric HPV:HIV VLPs were further characterized by HIV-1 anti-V3 and anti-2F5 monoclonal antibodies in vitro by using Western blot and indirect ELISA. Finally, the immunogenicity of HPV:HIV VLPs were assessed in BALB/c mice model. Because the development and manufacturing of an immunogenic HPV:HIV vaccine is still unachievable, this study provided a baseline strategy that may be worthy to support the global efforts to develop novel chimeric VLP-based vaccines for controlling HPV and HIV-1 infections
ca
dc.format.extent
206 p.
ca
dc.language.iso
eng
ca
dc.publisher
Universitat de Barcelona
dc.rights.license
L'accés als continguts d'aquesta tesi queda condicionat a l'acceptació de les condicions d'ús establertes per la següent llicència Creative Commons: http://creativecommons.org/licenses/by-nc/4.0/
ca
dc.rights.uri
http://creativecommons.org/licenses/by-nc/4.0/
*
dc.source
TDX (Tesis Doctorals en Xarxa)
dc.subject
Cèl·lules
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dc.subject
Células
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dc.subject
Cells
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dc.subject
Cromatografia
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dc.subject
Cromatografía
ca
dc.subject
Chromatography
ca
dc.subject
Vacunes
ca
dc.subject
Vacunas
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dc.subject
Vaccines
ca
dc.subject.other
Ciències de la Salut
ca
dc.title
Pre-clinical development of chimeric virus-like particles based HPV: HIV vaccines by using mammalian cell expression system: limitations and challenges
ca
dc.type
info:eu-repo/semantics/doctoralThesis
dc.type
info:eu-repo/semantics/publishedVersion
dc.subject.udc
576
ca
dc.contributor.director
Joseph Munné, Joan
dc.embargo.terms
cap
ca
dc.rights.accessLevel
info:eu-repo/semantics/openAccess


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