Summary
The determination of protein patterns in nasal secretions of healthy subjects can help in the early diagnosis of diseases such as acute sinusitis. The comparison of nasal lavage fluid collected from subjects with acute sinusitis before and after pharmacological treatment gives information about the drug effects on glandular secretions. Nasal secretions were stimulated with 1× NS (0.9% Normal Saline) and 24× NS in healthy subjects and in sinusitis subjects before and after pharmacological treatment. The nasal lavage fluid (NLF) proteins are precipitated with a solution of “acid-ethanol.” Using this solution, the high molecular weight proteins precipitate and separate from the low molecular weight proteins. The proteins are digested and the peptides are separated using a capillary liquid chromatographic system. Eluted peptides are analyzed on ESI-Q-TOF mass spectrometry instrument.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Baraniuk, J. N. (2000)Immunology and Allergy Clinics of North America (Lasley, M. and V. Altman, L. C. eds.), Saunders, Philadelphia, pp. 245–64.
Baraniuk, J. N., Staevska, M. (2004) Current Therapy in Allergy Immunology and Rheumatology (Lichtenstein, L. M., Busse, W. W. and Geha, R. S., eds.), Mosby, Philadelphia, pp. 17–24.
Meyer, P.,Andersson, M.,Persson, C.G., and Greiff, L. (2003) Steroid-sensitive indices of airway inflammation in children with seasonal allergic rhinitis. Pediatr. Allergy Immunol. 14, 60–65
Dahl, R., and Mygind, N. (1998) Mechanisms of airflow limitation in the nose and lungs. Clin. Exp. Allergy 28, 17–25.
Svensson, C., Andersson, M.,Greiff, L., and Persson, C.G. (1998) Nasal mucosal end organ hyperresponsiveness. Am. J. Rhinol. 12, 37–43.
Baraniuk, J. N., Ali, M.,Yuta, A., Fang, S-Y., and Naranch, K. (1999) Hypertonic saline nasal provocation stimulates nociceptive nerves, substance P release, and glandular mucus exocytosis in normal humans. Am. J. Respir. Crit. Care Med. 160, 655–62.
Sanico, A. M., Philip, G., Lai, G. K., and Togias, A. (1999) Hyperosmolar saline induces reflex nasal secretions, evincing neural hyperresponsiveness in allergic rhinitis. J. Appl. Physiol. 86, 1202–10.
Baraniuk, J. N., Clauw, D. J., and Gaumond, E. (1998) Rhinitis symptoms in chronic fatigue syndrome. Ann. Allergy Asthma Immunol. 81, 359–65.
Fukuda, K., Straus, S. E., Hickei, I., Sharpe, M. C., Dobbins, J. C., and Komaroff, A. (1994) The chronic fatigue syndrome: a comprehensive approach to its definition and study. Ann. Intern. Med. 121, 953–9.
Tabaton, M., Monaco, S., Cordone, M. P., Colucci, M., Giaccone, G., Tagliavini, F., and Zanusso, G. (2004) Prion deposition in olfactory biopsy of sporadic Creutzfeldt-Jakob disease. Ann Neurol. 55, 294–6.
Sloat, B. R., and Cui, Z. (2005) Strong mucosal and systemic immunities induced by nasal immunization with anthrax protective antigen protein incorporated in liposome-protamine-dna particles. Pharm Res. it. Dec 6; [Epub ahead of print]
Lindahl, M., Stahlbom, B., and Tagesson, C. (1995) Two-dimensional gel electrophoresis of nasal and onchoalveolar lavage fluids after occupational exposure. Electrophoresis 16, 1199–1204.
Lindahl, M., Stahlbom, B., Svartz, J., and Tagesson, C. (1998) Protein patterns of human nasal and bronchoalveolar lavage fluids analyzed with two-dimensional gel electrophoresis. Electrophoresis 19, 3222–29.
Lindahl, M., Stahlbom, B., and Tagesson, C. (1999) Newly identified proteins in human nasal and bronchoalveolar lavage fluids: potential biomedical and clinical applications. Electrophoresis 20, 3670–76.
Lindahl, M., Svartz, J., and Tagesson, C. (1999) Demonstration of different forms of the anti-inflammatory proteins lipocortin-1 and Clara cell protein-16 in human nasal and bronchoalveolar lavage fluids. Electrophoresis 20, 881–90.
Ghafouri, B.,Stahlbom, B., Tagesson, C., and Lindahl, M. (2002) Newly identified proteins in human nasal lavage fluid from non-smokers and smokers using two-dimensional gel electrophoresis and peptide mass fingerprinting. Proteomics 2, 112–20.
Lindahl, M., Stahlbom, B., andTagesson, C. (2001) Identification of a new potential airway irritation marker, palate lung nasal epithelial clone protein, in human nasal lavage fluid with two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight. Electrophoresis 22, 1795–1800.
Lindahl, M., Irander, K., Tagesson, C., and Stahlbom, B. (2004) Nasal lavage fluid and proteomics as means to identify the effects of the irritating epoxy chemical dimethylbenzylamine. Biomarkers 9, 56–70.
Casado, B., Pannell, L. K., Viglio, S., Iadarola, P. et al. (2004) Analysis of the sinusitis nasal lavage fluid proteome using capillary liquid chromatography interfaced to electrospray ionization-quadrupole time of flight- tandem mass spectrometry. Electrophoresis 25, 1386–93.
Casado, B., Pannell, L. K., Iadarola, P.,Baraniuk, J.N. (2005) Identification of human nasal mucous proteins using proteomics. Proteomics 5, 2949–59.
Kristiansson, M. H., Lindh, C. H., and Jonsson, B. A. (2003) Determination of hexahydrophthalic anhydride adducts to human serum albumin. Biomarkers 8, 343–59
Kristiansson, M. H., Lindh, C. H., andJonsson, B. A. (2004) Correlations between air levels of hexahydrophthalic anhydride (HHPA) and HHPA-adducted albumin tryptic peptides in nasal lavage fluid from experimentally exposed volunteers. Rapid Commun Mass Spectrom. 18, 1592–8.
Ali, M., Maniscalco, J., and Baraniuk, J. N. (1996) Spontaneous release of submucosal gland serous and mucous cell macromolecules from human nasal explants in vitro. Am. J. Physiol. 270, L595–L600.
Baraniuk, J. N., Silver, P. B., Kaliner, M. A., and Barnes, P. J. (1994) Int. Arch. Allergy Immunol. 103, 202–8.
Baraniuk, J. N., Okayama, M., Lundgren, J. D., Mullol, M. et al. (1990) Vasoactive intestinal peptide in human nasal mucosa. J. Clin. Invest. 86, 825–31.
Wu, H. C. H., Huang, H., Yeh, Lai-Su, L.,Barker, C. W. (2003) Protein family classification and functional annotation. Comput. Biol. Chem. 27, 37–47.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Casado, B., Iadarola, P., Pannell, L.K. (2008). Preparation of Nasal Secretions for Proteome Analysis. In: Posch, A. (eds) 2D PAGE: Sample Preparation and Fractionation. Methods in Molecular Biology™, vol 425. Humana Press. https://doi.org/10.1007/978-1-60327-210-0_7
Download citation
DOI: https://doi.org/10.1007/978-1-60327-210-0_7
Publisher Name: Humana Press
Print ISBN: 978-1-60327-209-4
Online ISBN: 978-1-60327-210-0
eBook Packages: Springer Protocols