Absolute quantification of norovirus capsid protein in food, water, and soil using synthetic peptides with electrospray and MALDI mass spectrometry

Erica M. Hartmann, David R. Colquhoun, Kellogg J. Schwab, Rolf U. Halden*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Norovirus infections are one of the most prominent public health problems of microbial origin in the U.S. and other industrialized countries. Surveillance is necessary to prevent secondary infection, confirm successful cleanup after outbreaks, and track the causative agent. Quantitative mass spectrometry, based on absolute quantitation with stable-isotope labeled peptides, is a promising tool for norovirus monitoring because of its speed, sensitivity, and robustness in the face of environmental inhibitors. In the current study, we present two new methods for the detection of the norovirus genogroup I capsid protein using electrospray and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The peptide TLDPIEVPLEDVR was used to quantify norovirus-like particles down to 500 attomoles with electrospray and 100 attomoles with MALDI. With MALDI, we also demonstrate a detection limit of 1 femtomole and a quantitative dynamic range of 5 orders of magnitude in the presence of an environmental matrix effect. Due to the rapid processing time and applicability to a wide range of environmental sample types (bacterial lysate, produce, milk, soil, and groundwater), mass spectrometry-based absolute quantitation has a strong potential for use in public health and environmental sciences.

Original languageEnglish (US)
Pages (from-to)525-532
Number of pages8
JournalJournal of Hazardous Materials
Volume286
DOIs
StatePublished - Apr 9 2015

Funding

We thank Mary Estes for providing the recombinant Norwalk virus virus-like particles and the Mass Spectrometry and Proteomics Facility at the Johns Hopkins Medical Institutions, Bob O'Meally, and Robert Cole for their assistance with experimental design and method development. This project was supported in part by Award Numbers R01ES015445 and R01ES020889 from the National Institute of Environmental Health Sciences (NIEHS). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIEHS or the National Institutes of Health (NIH). EMH was supported by a fellowship from the Arizona State University Security & Defense Systems Initiative.

Keywords

  • Absolute quantification
  • Environmental detection
  • Mass spectrometry
  • Norovirus

ASJC Scopus subject areas

  • Pollution
  • Waste Management and Disposal
  • Health, Toxicology and Mutagenesis
  • Environmental Engineering
  • Environmental Chemistry

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