Yeast-derived virus-like particles as components of novel vaccine prototypes
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Latvijas Universitāte
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eng
Abstract
Vīrusiem līdzīgās daļiņas (VLPs), tiek izmantotas gan fundamentālajā, gan pielietojamā
zinātnē, bet galvenokārt jaunu vakcīnu izveidē.
Šajā darbā raugu ekspresijas sistēma tika pielāgota (i) RNS bakteriofāgu VLP un (ii) HBc
veidoto VLP iegūšanai. Tika pārbaudīta septiņu dažādu fāgu VLP veidotspēja raugos
Saccharomyces cerevisiae un Pichia pastoris, respektīvi, Qβ, SP, fr, GA, AP205, PP7 un
fāgam φCb5.
Fermentācijas apstākļos tika panākta efektīva HBc VLP produkcija raugā P. pastoris, un
tika izstrādāta efektīva attīrīšanas metode.
Laboratorijas praksē tika ieviestas vēl divas raugu ekspresijas sistēmas:
Kluyveromyces lactis un Hansenula polymorpha. Tika konstruēta virkne jaunu vektoru
ekspresijai H. polymorpha.
Virus-like particles (VLPs) have been used for a wide range of applications, but especially in vaccine development. Yeast expression system was adapted for generation of (i) the bacteriophage VLPs (CPs) (ii) hepatitis B virus (HBV) core (nucleocapsid) particles. Seven different phages were tested for their CP gene expression, namely, Qβ, SP, fr, GA, AP205, PP7 and φCb5. High-level fermentor production was established for HBc VLPs in P. pastoris and purification scheme was developed. Yeast expression systems have been introduced in laboratory practice, namely, Kluyveromyces lactis and Hansenula polymorpha. A set of new vectors for expression in yeast H. polymorpha was constructed.
Virus-like particles (VLPs) have been used for a wide range of applications, but especially in vaccine development. Yeast expression system was adapted for generation of (i) the bacteriophage VLPs (CPs) (ii) hepatitis B virus (HBV) core (nucleocapsid) particles. Seven different phages were tested for their CP gene expression, namely, Qβ, SP, fr, GA, AP205, PP7 and φCb5. High-level fermentor production was established for HBc VLPs in P. pastoris and purification scheme was developed. Yeast expression systems have been introduced in laboratory practice, namely, Kluyveromyces lactis and Hansenula polymorpha. A set of new vectors for expression in yeast H. polymorpha was constructed.