Extensive fractionation and identification of proteins within nasal lavage fluids from allergic rhinitis and asthmatic chronic rhinosinusitis patients

Linda M. Benson, Christopher J. Mason, Oren Friedman, Hirohito Kita, Harold Robert (Bob) III Bergen, Douglas A. Plager

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Allergic rhinitis (AR), chronic rhinosinusitis (CRS), and asthma are prevalent airway diseases that can have a substantial impact on a patient's quality of life. MS analyses of biological fluids can effectively screen for proteins associated with disease processes, however, initial detection of diagnostic proteins is difficult due to protein complexity and dynamic range. To enhance the detection of lower abundance proteins, intact nasal lavage fluid (NLF) proteins from nonpolypoid AR and from asthmatic CRS patients were extensively fractionated prior to LC/MS/MS analysis. Pooled NLF samples were processed to remove low molecular weight molecules and high abundance plasma proteins. Anion exchange (AX) chromatography followed by RP-LC further separated the remaining intact NLF proteins. The resulting fractions were digested with trypsin and the peptides analyzed by LC/MS/ MS. Spectra were searched with MASCOT, SEQUEST, and X!Tandem to obtain peptide identifications and subsequently analyzed by Scaffold software to identify parent proteins with at least 99% confidence. The 197 identified proteins are compared to those previously cited in the literature and the workflow evaluated to determine the usefulness for the detection of lower abundance proteins. This is the first extensive list of NLF proteins generated from CRS patients with coexisting asthma.

Original languageEnglish (US)
Pages (from-to)44-56
Number of pages13
JournalJournal of Separation Science
Volume32
Issue number1
DOIs
StatePublished - Jan 2009

Fingerprint

Fractionation
Proteins
Fluids
Peptides
Chromatography
Scaffolds
Scaffolds (biology)
Trypsin
Anions
Blood Proteins
Ion exchange
Molecular weight
Negative ions
Molecules
Plasmas

Keywords

  • Allergic rhinitis
  • Chronic rhinosinusitis
  • Nasal lavage
  • Protein fractionation
  • Proteomics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

Cite this

Extensive fractionation and identification of proteins within nasal lavage fluids from allergic rhinitis and asthmatic chronic rhinosinusitis patients. / Benson, Linda M.; Mason, Christopher J.; Friedman, Oren; Kita, Hirohito; Bergen, Harold Robert (Bob) III; Plager, Douglas A.

In: Journal of Separation Science, Vol. 32, No. 1, 01.2009, p. 44-56.

Research output: Contribution to journalArticle

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