https://scholars.lib.ntu.edu.tw/handle/123456789/634035
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.author | Chien, Man Ling | en_US |
dc.contributor.author | Yu, Chun Fu | en_US |
dc.contributor.author | CHING-TSAN HUANG | en_US |
dc.date.accessioned | 2023-07-24T02:22:43Z | - |
dc.date.available | 2023-07-24T02:22:43Z | - |
dc.date.issued | 2023-06-01 | - |
dc.identifier.issn | 2311-5637 | - |
dc.identifier.uri | https://scholars.lib.ntu.edu.tw/handle/123456789/634035 | - |
dc.description.abstract | Many efforts in norovirus vaccine development have focused on subunit or recombinant protein vaccines, such as subviral P particles formed by the protruding (P) domain of VP1. P particles are immunogenic and have a region with a human histo-blood group antigen binding site, an interaction critical for infecting the host. In the past, only intracellular NoV P proteins expressed in Escherichia coli and Pichia pastoris were reported, and the low yield and difficulty in purification limited their applications. In this study, the Taiwan-native NoV P domain was successfully expressed and secreted by P. pastoris. The secretion efficiency was greatly enhanced by integrating oligosaccharyl transferase (Ost1) into the α-factor signal peptide and coexpressing Hac1. The production of NoV P in fermentation cultures reached 345 mg/L, and the purity and recovery were 94.8% and 66.9%, respectively, after only ion-exchange chromatography. Transmission electron microscopy analysis showed that the small P particles were mostly ring-, square-, and triangle-shaped, with diameters of 10-15 nm. The biological activity of NoV P was confirmed by saliva-binding assay using human histo-blood group antigen. This study describes the secretion and characterization of the Taiwan-native norovirus P domain in P. pastoris. Particles formed from the P domain were similar in size, morphology, and binding ability to those expressed intracellularly. The strategy described in this study provides great potential in scale-up production and antiviral vaccine development. | en_US |
dc.language.iso | en | en_US |
dc.publisher | MDPI | en_US |
dc.relation.ispartof | Fermentation | en_US |
dc.subject | Hac1 | norovirus | oligosaccharyl transferase 1 (Ost1) | P particle | tag-free P protein | en_US |
dc.title | Extracellular Production of the Taiwan-Native Norovirus P Domain Overexpressed in Pichia pastoris | en_US |
dc.type | journal article | en |
dc.identifier.doi | 10.3390/fermentation9060498 | - |
dc.identifier.scopus | 2-s2.0-85163364911 | - |
dc.identifier.url | https://api.elsevier.com/content/abstract/scopus_id/85163364911 | - |
dc.relation.pages | Article number 498 | en_US |
dc.relation.journalvolume | 9 | en_US |
dc.relation.journalissue | 6 | en_US |
item.fulltext | no fulltext | - |
item.languageiso639-1 | en | - |
item.openairecristype | http://purl.org/coar/resource_type/c_6501 | - |
item.cerifentitytype | Publications | - |
item.openairetype | journal article | - |
item.grantfulltext | none | - |
crisitem.author.dept | Biochemical Science and Technology | - |
crisitem.author.orcid | 0000-0003-1019-784X | - |
crisitem.author.parentorg | College of Life Science | - |
顯示於: | 生化科技學系 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。