Abstract
Bacterial surface proteins are often investigated as potential vaccine candidates and biomarkers of virulence. In this chapter, a novel method for identifying bacterial surface proteins is presented, which combines immunoproteomic with immunoserologic techniques. Immunoproteomics, involving the separation of proteins by two-dimensional electrophoresis (2-DE) and Western blotting, has become an increasingly popular method for identifying immunoreactive proteins. In conventional serological technique, cross-absorption is a powerful method used to minimize cross reaction during agglutination assays. The serum pre-absorption process in our method was developed from cross-absorption but modified to remove antibodies that recognized bacterial surface antigens, thereby generating pre-absorbed sera. These pre-absorbed sera were used in Western blotting after 2-DE to find bacterial surface protein antigens. This new method has been proven to be a useful tool for identifying surface proteins, and aid in the development of new vaccine subunits and disease diagnostics.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wolters DA, Washburn MP, Yates JR III (2001) An automated multidimensional protein identification technology for shotgun proteomics. Anal Chem 73:5683–5690
Zhu H, Pan S, Gu S, Bradbury EM, Chen X (2002) Amino acid residue specific stable isotope labeling for quantitative proteomics. Rapid Commun Mass Spectrom 16:2115–2123
Hempel K, Pane-Farre J, Otto A, Sievers S, Hecker M, Becher D (2010) Quantitative cell surface proteome profiling for SigB-dependent protein expression in the human pathogen Staphylococcus aureus via biotinylation approach. J Proteome Res 9:1579–1590
Tjalsma H, Lambooy L, Hermans PW, Swinkels DW (2008) Shedding & shaving: disclosure of proteomic expressions on a bacterial face. Proteomics 8:1415–1428
Cahill DJ (2001) Protein and antibody arrays and their medical applications. J Immunol Methods 250:81–91
Mittal KR, Higgins R, Lariviere S (1982) Evaluation of slide agglutination and ring precipitation tests for capsular serotyping of Haemophilus pleuropneumoniae. J Clin Microbiol 15:1019–1023
Gottschalk M, Segura M (2000) The pathogenesis of the meningitis caused by Streptococcus suis: the unresolved questions. Vet Microbiol 76:259–272
Zhang W, Liu G, Tang F, Shao J, Lu Y, Bao Y, Yao H, Lu C (2011) Pre-absorbed immunoproteomics: a novel method for the detection of Streptococcus suis surface proteins. PLoS One 6:e21234
Winterhoff N, Goethe R, Gruening P, Rohde M, Kalisz H, Smith HE, Valentin-Weigand P (2002) Identification and characterization of two temperature-induced surface-associated proteins of Streptococcus suis with high homologies to members of the Arginine Deiminase system of Streptococcus pyogenes. J Bacteriol 184:6768–6776
Robertson ID, Blackmore DK (1989) Occupational exposure to Streptococcus suis type 2. Epidemiol Infect 103:157–164
Kussmann M, Nordhoff E, Rahber-Nielsen H et al (1997) Matrix-assisted laser desorption/ionization mass spectrometry sample preparation techniques designed for various peptide and protein analytes. J Mass Spectrom 32:593–601
Ling E, Feldman G, Portnoi M, Dagan R, Overweg K, Mulholland F, Chalifa-Caspi V, Wells J, Mizrachi-Nebenzahl Y (2004) Glycolytic enzymes associated with the cell surface of Streptococcus pneumoniae are antigenic in humans and elicit protective immune responses in the mouse. Clin Exp Immunol 138:290–298
Sellman BR, Howell AP, Kelly-Boyd C, Baker SM (2005) Identification of immunogenic and serum binding proteins of Staphylococcus epidermidis. Infect Immun 73:6591–6600
Fernandez-Arenas E, Molero G, Nombela C, Diez-Orejas R, Gil C (2004) Low virulent strains of Candida albicans: unravelling the antigens for a future vaccine. Proteomics 4:3007–3020
Bunai K, Yamane K (2005) Effectiveness and limitation of two-dimensional gel electrophoresis in bacterial membrane protein proteomics and perspectives. J Chromatogr B Analyt Technol Biomed Life Sci 815:227–236
Siegel JL, Hurst SF, Liberman ES, Coleman SE, Bleiweis AS (1981) Mutanolysin-induced spheroplasts of Streptococcus mutants are true protoplasts. Infect Immun 31:808–815
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Liu, G., Zhang, W., Lu, C. (2013). Pre-absorbed Immunoproteomics: A Novel Method for the Detection of Bacterial Surface Proteins. In: Fulton, K., Twine, S. (eds) Immunoproteomics. Methods in Molecular Biology, vol 1061. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-589-7_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-589-7_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-588-0
Online ISBN: 978-1-62703-589-7
eBook Packages: Springer Protocols