The Virus

Ruis, Christopher; Thorne, Lucy; Breuer, Judith

doi:10.1007/978-3-030-27209-8_2

Cited by 2 publications

(2 citation statements)

References 175 publications

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“…Noroviruses (previously known as Norwalk or Norwalk-like viruses) are non-enveloped, singlestranded RNA viruses 1 known to cause a disease initially described in 1929 as the 'winter vomiting disease'. 2 It is the most common cause of viral gastroenteritis in humans with about 685 million cases worldwide, 200 000 deaths per year, and one in five cases being acute.…”

Section: Introductionmentioning

confidence: 99%

An Isotope Dilution Mass Spectrometry Assay to track Norovirus-Like Particles in Vaccine Process Intermediates by Quantifying Capsid Protein VP1

Jhingree

Boisvert

Mercier

2023

Preprint

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The coronavirus disease (COVID-19) pandemic shows the rapid pace at which vaccine development can occur which highlights the need for more fast and efficient analytical methodologies to track and characterize candidate vaccines during manufacturing and purification processes. The candidate vaccine in this work comprises plant-derived Norovirus-like particles (NVLPs) which are structures that mimic the virus but lack any infectious genetic material. Presented here is a liquid chromatography-tandem mass spectrometry (LC-MS/MS) methodology for the quantification of viral protein VP1, the main component of the NVLPs in this study. It combines isotope dilution mass spectrometry (IDMS) with multiple reaction monitoring (MRM) to quantify targeted peptides in process intermediates. Multiple MRM transitions (precursor/product ion pairs) for VP1 peptides were tested with varying MS source conditions and collision energies. Final parameter selection for quantification includes three peptides with two MRM transitions each offering maximum detection sensitivity under optimized MS conditions. For quantification, a known concentration of the isotopically labeled version of the peptides to be quantified was added into working standard solutions to serve as an internal standard (IS); calibration curves were generated for concentration of native peptide vs. the peak area ratio of native-to-isotope labeled peptide. VP1 peptides in samples were quantified with labeled versions of the peptides added at the same level as that of the standards. Peptides were quantified with limit of detection (LOD) as low as 1.0 fmol/μL and limit of quantitation (LOQ) as low as 2.5 fmol/μL. NVLP preparations spiked with known quantities of either native peptides or drug substance (DS) comprising assembled NVLPs produced recoveries indicative of minimal matrix effects. Overall, we report a fast, specific, selective, and sensitive LC-MS/MS strategy to track NVLPs through the purification steps of the DS of a Norovirus candidate vaccine.

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Section: Introductionmentioning

confidence: 99%

An Isotope Dilution Mass Spectrometry Assay to track Norovirus-Like Particles in Vaccine Process Intermediates by Quantifying Capsid Protein VP1

Jhingree

Boisvert

Mercier

2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Noroviruses (previously known as Norwalk or Norwalk-like viruses) are non-enveloped, single-stranded RNA viruses 1 known to cause a disease initially described in 1929 as the ‘winter vomiting disease’. 2 It is the most common cause of viral gastroenteritis in humans with about 685 million cases worldwide, 200 000 deaths per year, and one in five cases being acute.…”

Section: Introductionmentioning

confidence: 99%