Generation of menangle virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae

Juozapaitis, Mindaugas; Serva, Andrius; Kucinskaite, Indre; Žvirblienė, Aurelija; Slibinskas, Rimantas; Staniulis, J.; Sasnauskas, Kęstutis; Shiell, Brian J.; Bowden, Timothy R.; Michalski, Wojtek P.

doi:10.1016/j.jbiotec.2007.05.013

Cited by 13 publications

(5 citation statements)

References 39 publications

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“…Establishment, validation and comparison of a PUUVBawa N protein-based indirect IgG and capture ELISA Purified, yeast-expressed viral antigens have broadly been used for diagnostics of virus infections in humans and animals [76][77][78][79][80][81]. We have recently described indirect and mAb-capture IgG-ELISAs for the detection of human infections with SNV, ANDV, PUUV, DOBV and HTNV [32,[36][37][38].…”

Section: Reactivity Of Human Sera and Mabs With Different Puuv N Antimentioning

confidence: 99%

“…4 Western blot reactivity of the purified recombinant PUUVBawa N protein (aa 2-433) of about 49 kDa with His-tag specific mAbs 6E8 and 7B8 and mAbs raised against N proteins of PUUV (mAbs 5E11, 5C5, 2C6 [39], 5A3, 4C3, 1C12, A1C5 [42], 5E1, 2E12, [40,41], 7A5), Sin Nombre virus (SNV; mAb 5F1/F7), Andes virus (ANDV; mAb 5C2/E10), ANDV/SNV (mAb 4H3, 7G2 [44]) and Hantaan virus (HTNV; mAb E5/G6, Eco2 [43]; B5D9 [42]). The epitope localization of the PUUV-, HTNV and SNV/ANDV-reactive mAbs has been described in detail in [43,78,79]. Epitope specificity for mAbs 5C2/E10 and 5F1/F7 is given according to the information of the manufacturer (Immunological and Biochemical Testsystems GmbH, Reutlingen, Germany) Germany one sample was identified as false negative (Table 5).…”

Section: Reactivity Of Human Sera and Mabs With Different Puuv N Antimentioning

confidence: 99%

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Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

et al. 2011

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Puumala virus (PUUV) is the predominant hantavirus species in Germany causing large numbers of mild to moderate cases of haemorrhagic fever with renal syndrome (HFRS). During an outbreak in South-East Germany in 2004 a novel PUUV subtype designated Bavaria was identified as the causative agent of HFRS in humans [1]. Here we present a molecular characterization of this PUUV strain by investigating novel partial and almost entire nucleocapsid (N) protein-encoding small (S-) segment sequences and partial medium (M-) segment sequences from bank voles (Myodes glareolus) trapped in Lower Bavaria during 2004 and 2005. Phylogenetic analyses confirmed their classification as subtype Bavaria, which is further subdivided into four geographical clusters. The entire N protein, harbouring an amino-terminal hexahistidine tag, of the Bavarian strain was produced in yeast Saccharomyces cerevisiae and showed a slightly different reactivity with N-specific monoclonal antibodies, compared to the yeast-expressed N protein of the PUUV strain Vranica/Hällnäs. Endpoint titration of human sera from different parts of Germany and from Finland revealed only very slight differences in the diagnostic value of the different recombinant proteins. Based on the novel N antigen indirect and monoclonal antibody capture IgG-ELISAs were established. By using serum panels from Germany and Finland their validation demonstrated a high sensitivity and specificity. In summary, our investigations demonstrated the Bavarian PUUV strain to be genetically divergent from other PUUV strains and the potential of its N protein for diagnostic applications.

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Section: Reactivity Of Human Sera and Mabs With Different Puuv N Antimentioning

confidence: 99%

Section: Reactivity Of Human Sera and Mabs With Different Puuv N Antimentioning

confidence: 99%

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

et al. 2011

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“…Therefore, the icosahedral VLP platform remains a good option for many applications due to the advantages of purification as well as a clear regulatory pathway due to the availability of licensed vaccines. The introduction of yeast expression systems into the production of complex VLPs can potentially improve the safety and quality control of biotechnological processes [85,86,87,88,89,90,91]. …”

Section: Discussionmentioning

confidence: 99%

Development of Virus-Like Particle Technology from Small Highly Symmetric to Large Complex Virus-Like Particle Structures

2013

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Virus-like particle (VLP) technology is a promising approach for the construction of novel vaccines, diagnostic tools, and gene therapy vectors. Initially, VLPs were primarily derived from non-enveloped icosahedral or helical viruses and proved to be viable vaccine candidates due to their effective presentation of epitopes in a native conformation. VLP technology has also been used to prepare chimeric VLPs decorated with genetically fused or chemically coupled epitope stretches selected from immunologically defined target proteins. However, structural constraints associated with the rigid geometrical architecture of icosahedral or helical VLPs pose challenges for the expression and presentation of large epitopes. Complex VLPs derived from non-symmetric enveloped viruses are increasingly being used to incorporate large epitopes and even full-length foreign proteins. Pleomorphic VLPs derived from influenza or other enveloped viruses can accommodate multiple full-length and/or chimeric proteins that can be rationally designed for multifunctional purposes, including multivalent vaccines. Therefore, a second generation of VLP carriers is represented by complex particles reconstructed from natural or chimeric structural proteins derived from complex enveloped viruses. Further development of safe and efficient VLP nanotechnology may require a rational combination of both approaches.

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“…Following retrieval, the sections were quenched with hydrogen peroxide. Sections were then incubated for 1 h with a rabbit serum, ‘372’ (Petraityte et al , 2009), directed against CsCl gradient-purified, recombinant MenPV nucleocapsid protein, which was expressed in yeast (Juozapaitis et al , 2007). This was followed by HPR-conjugated secondary antibody (Envision Flex+; Dako).…”

Section: Methodsmentioning

confidence: 99%

Menangle virus, a pteropid bat paramyxovirus infectious for pigs and humans, exhibits tropism for secondary lymphoid organs and intestinal epithelium in weaned pigs

Bowden

Bingham

Harper

et al. 2012

Journal of General Virology

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This study is the first report of experimental infection and transmission of Menangle virus (MenPV) in pigs. Isolated in 1997 from piglets that were stillborn at a large commercial piggery in New South Wales, Australia, MenPV is a recently identified paramyxovirus of bat origin that causes severe reproductive disease in pigs and an influenza-like illness, with a rash, in humans. Although successfully eradicated from the infected piggery, the virus was only isolated from affected fetuses and stillborn piglets during the period of reproductive disease, and thus the mode of transmission between pigs was not established. To investigate the pathogenesis of MenPV, we undertook time-course studies in 6-week-old pigs following intranasal administration of a low-passage, non-plaque-purified isolate from the lung of an infected stillborn piglet. Viraemia was of short duration and low titre, as determined by real-time RT-PCR and virus isolation. Following an incubation period of 2–3 days, virus was shed in nasal and oral secretions, faeces and urine, typically for less than 1 week. Cessation of shedding correlated with the development of neutralizing antibodies in sera. Secondary lymphoid organs and intestine were identified, using quantitative real-time RT-PCR, as major sites of viral replication and dissemination, and this was confirmed by positive immunolabelling of viral antigen within various lymphoid tissues and intestinal epithelium. These data provide new insights into the pathogenesis of MenPV in weaned pigs, and will facilitate future control and eradication programmes should it ever re-emerge in the pig population.

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Generation of menangle virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae

Cited by 13 publications

References 39 publications

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein

Development of Virus-Like Particle Technology from Small Highly Symmetric to Large Complex Virus-Like Particle Structures

Menangle virus, a pteropid bat paramyxovirus infectious for pigs and humans, exhibits tropism for secondary lymphoid organs and intestinal epithelium in weaned pigs

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