Abstract
The analysis of proteins and peptides in biological fluids is becoming more important as they are potential sources of diagnostic biomarkers of disease. The complexity of body fluids is such that no single technique can both identify and quantify all the constituents present. It therefore requires pre-concentration of the compounds of interest and hyphenated techniques like capillary electrophoresis (CE) or high-performance liquid chromatography (HPLC) coupled to a mass spectrometer (MS) to provide the separation, identification, and quantification of the proteins and peptides under investigation. Here we provide protocols for sample preparation of various body fluids, namely, urine, plasma, and cerebrospinal fluid, and how to obtain maximum sensitivity and selectivity of the small proteins and peptides under investigation by CE-MS.
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
Fliser D, Novak J, Thongboonkerd V, Argiles A, Jankowski V, Girolami M, Jankowski J, Mischak H (2007) Advances in urinary proteome analysis and biomarker discovery. J Am Soc Nephrol 18:1057–1071
Decramer S, de Gonzalez PA, Breuil B, Mischak H, Monsarrat B, Bascands JL, Schanstra JP (2008) Urine in clinical proteomics. Mol Cell Proteomics 7:1850–1862
Mischak H, Kolch W, Aivalotis M, Bouyssie D, Court M, Dihazi H, Dihazi GH, Franke J, Garin J, Gonzales de Peredo A, Iphöfer A, Jansch L, Lacroix C, Makridakis M, Masselon C, Metzger J, Monsarrat B, Mrug M, Norling M, Novak J, Pich A, Pitt A, Bongcam-Rudloff E, Siwy J, Suzuki H, Thongboonkerd V, Wang L, Zoidakis J, Zurbig P, Schanstra J, Vlahou A (2010) Comprehensive human urine standards for comparability and standardization in clinical proteome analysis. Proteomics Clin Appl 4:464–478
Zürbig P, Renfrow MB, Schiffer E, Novak J, Walden M, Wittke S, Just I, Pelzing M, Neususs C, Theodorescu D, Root C, Ross M, Mischak H (2006) Biomarker discovery by CE-MS enables sequence analysis via MS/MS with platform-independent separation. Electrophoresis 27:2111–2125
Tomer KB (2001) Separations Combined with Mass Spectrometry. Chem Rev 101:297–328
Neususs C, Pelzing M, Macht M (2002) A robust approach for the analysis of peptides in the low femtomole range by capillary electrophoresis-tandem mass spectrometry. Electrophoresis 23:3149–3159
Schmitt-Kopplin P, Frommberger M (2003) Capillary electrophoresis-mass spectrometry: 15 years of developments and applications. Electrophoresis 24:3837–3867
Tu C, Rudnick PA, Martinez MY, Cheek KL, Stein SE, Slebos RJ, Liebler DC (2010) Depletion of abundant plasma proteins and limitations of plasma proteomics. J Proteome Res 9:4982–4991
Roche S, Gabelle A, Lehmann S (2009) Clinical proteomics of the cerebrospinal fluid: towards the discovery of new biomarkers. Proteomics Clin Appl 2:428–436
Weissinger EM, Wittke S, Kaiser T, Haller H, Bartel S, Krebs R, Golovko I, Rupprecht HD, Haubitz M, Hecker H, Mischak H, Fliser D (2004) Proteomic patterns established with capillary electrophoresis and mass spectrometry for diagnostic purposes. Kidney Int 65:2426–2434
Jantos-Siwy J, Schiffer E, Brand K, Schumann G, Rossing K, Delles C, Mischak H, Metzger J (2009) Quantitative Urinary Proteome Analysis for Biomarker Evaluation in Chronic Kidney Disease. J Proteome Res 8:268–281
Lewczuk P, Kornhuber J, Wiltfang J (2006) The German competence net dementias: standard operating procedures for the neurochemical dementia diagnostics. J Neural Transm 113:1075–1080
Cham BE, Knowles BR (1976) A solvent system for delipidation of plasma or serum without protein precipitation J. Lipid Res 17:176–181
Neuhoff N, Kaiser T, Wittke S, Krebs R, Pitt A, Burchard A, Sundmacher A, Schlegelberger B, Kolch W, Mischak H (2004) Mass spectrometry for the detection of differentially expressed proteins: a comparison of surface-enhanced laser desorption/ionization and capillary electrophoresis/mass spectrometry. Rapid Commun Mass Spectrom 18:149–156
Theodorescu D, Wittke S, Ross MM, Walden M, Conaway M, Just I, Mischak H, Frierson HF (2006) Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. Lancet Oncol 7:230–240
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Albalat, A., Bitsika, V., Zurbig, P., Siwy, J., Mullen, W. (2013). High-Resolution Proteome/Peptidome Analysis of Body Fluids by Capillary Electrophoresis Coupled with MS. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_12
Download citation
DOI: https://doi.org/10.1007/978-1-62703-296-4_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-295-7
Online ISBN: 978-1-62703-296-4
eBook Packages: Springer Protocols