Abstract
Noroviruses cause immense sporadic gastroenteritis outbreaks worldwide. Emerging genotypes, which are divided based on the sequence of the major capsid protein VP1, further enhance this public threat. Self-assembling properties of the human norovirus major capsid protein VP1 are crucial for using virus-like particles (VLPs) for vaccine development. However, there is no vaccine available yet. Here, VLPs from different variants produced in insect cells were characterized in detail using a set of biophysical and structural tools. We used native mass spectrometry, gas-phase electrophoretic mobility molecular analysis, and proteomics to get clear insights into particle size, structure, and composition, as well as stability. Generally, noroviruses have been known to form mainly T = 3 particles. Importantly, we identified a major truncation in the capsid proteins as a likely cause for the formation of T = 1 particles. For vaccine development, particle production needs to be a reproducible, reliable process. Understanding the underlying processes in capsid size variation will help to produce particles of a defined capsid size presenting antigens consistent with intact virions. Next to vaccine production itself, this would be immensely beneficial for bio-/nano-technological approaches using viral particles as carriers or triggers for immunological reactions.
Highlights
A vast number of nonbacterial gastroenteritis cases worldwide is caused by human noroviruses [1]
In order to characterize hNoVLPs in detail and gain more insights into size determination, we extended our previous native mass spectrometry (MS) studies with a set of biophysical methods
Native mass spectra reveal the lack of T = 3 particles at neutral pH and moderate ionic strength (Figure 1)
Summary
A vast number of nonbacterial gastroenteritis cases worldwide is caused by human noroviruses (hNoVs) [1]. Norovirus infection especially poses an acute threat to children, immunocompromised individuals and elderly people. Already a small number of particles is sufficient for infection [2]. Gastroenteritis outbreaks happen worldwide with new hNoVvariants occurring sporadically. Human noroviruses are non-enveloped and a member of the Caliciviridae family. They have a positive sense, single strand, approx. 7.7 kb RNA genome organized into three open reading frames (ORFs) and a poly(A) tail. ORF1 encodes non-structural proteins, ORF2 the major capsid protein VP1, and ORF 3 the minor structural protein VP2 [3,4]
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