title: Improved preparation of nasal lavage fluid (NLF) as a noninvasive sample for proteomic biomarker discovery.
abstract: Nasal lavage fluid (NLF) becomes more and more important as a noninvasive patient sample serving as a new opportunity to discover new biomarkers in diverse human diseases comprising mainly respiratory disorders. This was supported by the observation that the protein profile of NLF differs from conventional samples of i.e. whole blood, hence being capable to complement or even expand the so far biomarker index. Since sample acquisition and processing are the most crucial steps for a profound and sensitive identification we present here a modified protocol of NLF generation and measurement. We show that mild washing steps for sample generation followed by column-mediated concentration and acetone precipitation are appropriate steps to minimize serum leakage by concomitantly highlighting proteins which represent typical components of NLF. This is shown by separation of proteins via 2D-PAGE followed by LC-MS/MS as well as Gel-LC-MS/MS measurements of cut and digested protein spots/bands.
  
  
  CONCLUSIONS
  For a better understanding of the molecular mechanisms underlying respiratory diseases NLF samples are favored sources for protein research. NLF acquisition and sample processing were impaired so far by the problem of blood serum leakage and high salt content. Here, we present a modified protocol of NLF generation leading to the display of typical inventory of NLF proteins combined with reduced salt and serum contaminants. By this, both assay reproducibility and the detection of up- or down-regulated proteins reliably can be discovered in the case of biomarker screenings in a disease versus control design. This article is part of a Special Issue entitled: Neuroproteomics: Applications in Neuroscience and Neurology.
authors: Abteilung für Neuroanatomie und Molekulare Hirnforschung, Medizinische Fakultaet, Ruhr-Universitaet Bochum, 44801 Bochum, Germany.
authors: Bodo
authors: B
authors: Schoenebeck
authors: Abteilung für Medizinische Proteomik/Bioanalytik, Medizinisches Proteom-Center, Ruhr-Universitaet Bochum, 44801 Bochum, Germany.
authors: Caroline
authors: C
authors: May
authors: Department of Otolaryngology, Phillips Universität, Marburg, Germany.
authors: Christian
authors: C
authors: Güldner
authors: Department of Neurology, Technische Universität München, Munich, Germany and German Center for Neurodegenerative Diseases (DZNE), Munich, Germany.
authors: Gesine
authors: G
authors: Respondek
authors: Paracelsus Elena-Klinik, 34128 Kassel, Germany.
authors: Brit
authors: B
authors: Mollenhauer
authors: Lehrstuhl für Translationale Neurodegeneration, Technische Universität München, Germany; Deutsches Zentrum für Neurodegenerative Erkrankungen, 81377 München, Germany.
authors: Guenter
authors: G
authors: Hoeglinger
authors: Leibniz-Institut für Analytische Wissenschaften -ISAS- e.V., Dortmund, Germany.
authors: Helmut E
authors: HE
authors: Meyer
authors: Abteilung für Funktionelle Proteomik, Medizinisches Proteom-Center, Ruhr-Universitaet Bochum, 44801 Bochum, Germany. Electronic address: Katrin.Marcus@rub.de.
authors: Katrin
authors: K
authors: Marcus
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002908:Chronic Disease; D005260:Female; D006801:Humans; D008297:Male; D017817:Nasal Lavage Fluid; D040901:Proteomics / Q000379:methods*; D006255:Rhinitis, Allergic, Seasonal / Q000378:metabolism*; D012852:Sinusitis / Q000378:metabolism*
chemical_list: D015415:Biomarkers
keywords: 2D-gel electrophoresis; Nasal lavage fluid