Abstract
Calcified aortic valve disease is a slowly progressive disorder that ranges from mild valve thickening with no obstruction of blood flow, known as aortic sclerosis, to severe calcification with impaired leaflet motion or aortic stenosis. Until now, aortic stenosis (AS) was thought to result from aging and “wear and tear” of the aortic valve, but nowadays, it is known that it presents the same risk factors as atherosclerosis and cardiovascular diseases.
A proteomic analysis of plasma could permit to identify the changes in protein expression induced by AS in this biological sample. However, the characterization of human plasma proteome is a very complicated task, due to the wide dynamic range of concentration that separates the most abundant proteins and the less common ones (10–12 orders of magnitude). For this reason, plasma analysis requires pre-fractionation methods, and several such techniques are currently used to deplete albumin and other abundant plasma proteins.
In this work we describe two different and optimized protocols to decrease the plasma proteome complexity for proteomic analysis. With this, comprehensive and systematic characterization of the plasma proteome in the healthy and diseased aortic stenosis (AS) state will greatly facilitate the development of “useful” biomarkers for early disease detection, clinical diagnosis, and therapy.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Iung B, Baron G, Butchart E, Delahaye F, Gohlke-Bärwolf C, Levang OW et al (2003) A prospective survey of patients with valvular heart disease in Europe: the Euro Heart Survey on Valvular Heart Disease. Eur Heart J 24:1231–1243
Lindroos M, Kupari M, Heikkila J, Tilvis R (1993) Prevalence of aortic valve abnormalities in the elderly: an echocardiographic study of a random population sample. J Am Coll Cardiol 21:1220–1225
Pomerance A (1972) Pathogenesis of aortic stenosis and its relation to age. Br Heart J 34:569–574
Otto CM, Kuusisto J, Reichenbach DD, Gown AM, O’Brien KD (1994) Characterization of the early lesion of `degenerative’ valvular aortic stenosis: histologic and immunohistochemical studies. Circulation 90:844–853
Rajamannan NM (2010) Mechanisms of aortic valve calcification: the LDL-density-radius theory: a translation from cell signalling to physiology. Am J Physiol Heart Circ Physiol 298:H5–H15
Veenstra TD, Conrads TP, Hood BL, Avellino AM, Ellenbogen RG, Morrison RS (2005) Biomarkers: mining the biofluid proteome. Mol Cell Proteomics 4:409–418
Anderson NL, Anderson NG (2002) The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 11:845–867
Anderson NL (2005) Candidate-based proteomics in the search for biomarkers of cardiovascular disease. J Physiol 563:23–60
Zolotarjova J, Martosella G, Nicol J, Bailey B, Boyes E, Barrett WC (2005) Differences among techniques for high abundant protein depletion. Proteomics 5:3304–3313
Wang YY, Cheng P, Chan DW (2003) A simple affinity spin tube filter method for removing high-abundant common proteins or enriching low-abundant biomarkers for serum proteomic analysis. Proteomics 3:243–248
Travis J, Bowen J, Tewksbury D, Johnson D, Pannell R (1976) Isolation of albumin from whole human plasma and fractionation of albumin-depleted plasma. Biochem J 157:301–306
Travis J, Pannell R (1973) Selective removal of albumin from plasma by affinity chromatography. Clin Chim Acta 49:49–52
Leatherbarrow RJ, Dean PD (1980) Studies on the mechanism of binding of serum albumin to immobilized Cibacron Blue F3GA. Biochem J 189:27–34
Seam N, Gonzales DA, Kern SJ, Hortin GL, Hoehn GT, Suffredini AF (2007) Quality control of serum albumin depletion for proteomic analysis. Clin Chem 53:1915–1920
Granger J, Siddiqui J, Copeland S, Remick D (2005) Albumin depletion of human plasma also removes low abundance proteins including the cytokines. Proteomics 5:4713–4718
Righetti PG, Boschetti E, Lomas L, Citterio A (2006) Protein equalizer technology: the quest for a “democratic proteome”. Proteomics 6:3980–3992
Sihlbom C, Kanmert I, von Bahr H, Davidsson P (2008) Evaluation of the combination of bead technology with SELDI-TOF-MS and 2-D DIGE for detection of plasma proteins. J Proteome Res 9:4191–4198
Paulus A, Freeby S, Academia K, Thulassiraman V (2009) Accessing low-abundance proteins in serum and plasma with a novel, simple enrichment and depletion method. Tech note 5632. Sample preparation. BioRad, Life science Group 6000. James Watson Drive, Hercules, CA.
Bjorhall K, Miliotis T, Davidsson P (2005) Comparison of different depletion strategies for improved resolution in proteomic analysis of human serum samples. Proteomics 5:307–317
Pieper R, Su Q, Gatlin CL, Huang ST, Anderson NL, Steiner S (2003) Multi-component immunoaffinity subtraction chromatography: an innovative step towards a comprehensive survey of the human plasma proteome. Proteomics 3:422–432
Gil-Dones F, Darde VM, Alonso Orgaz S, Lopez-Almodovar LF, Mourino-Alvarez L, Padial LR, Vivanco F, Barderas MG (2012) Inside human aortic stenosis: a proteomic analysis of plasma. J Proteomics 75:1639–1653
Gianazza E, Arnaud P (1982) A general method for fractionation of plasma proteins. Dye-ligand affinity chromatography on immobilized Cibacron blue F3-GA. Biochem J 201:129–136
Greenough C, Jenkins RE, Kiterringham NR, Pirmohamed M, Park BK, Pennington SR (2004) A method for the rapid depletion of albumin and immunoglobulin from human plasma. Proteomics 4:3107–3111
Acknowledgements
We thank the Proteomics Unit of the HNP for assistance and Carmen Bermudez for her technical support. This work was supported by grants from the Instituto de Salud Carlos III (FIS PI070537, PI11/02239), grants from Fondos Feder-Redes temáticas de Investigación Cooperativa en Salud (RD06/0014/1015), and grants from Fundación para la Investigación Sanitaria de Castilla-La Mancha (FISCAM PI2008-08, PI2008-28, PI2008-52).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Gil-Dones, F., Darde, V.M., Vivanco, F., Barderas, M.G. (2013). A Comparative Study of Immunodepletion and Equalization Methods for Aortic Stenosis Human Plasma. In: Vivanco, F. (eds) Heart Proteomics. Methods in Molecular Biology, vol 1005. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-386-2_20
Download citation
DOI: https://doi.org/10.1007/978-1-62703-386-2_20
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-385-5
Online ISBN: 978-1-62703-386-2
eBook Packages: Springer Protocols