Abstract
The importance of serum/plasma as a source of clinically relevant biomarkers/surrogate markers of human disease has increased significantly over the last decade (1,2), and modern proteomic methods have evolved and been adapted to meet the demand. The specific challenges facing serum analysis include the wide dynamic range in the concentration of individual components and the tremendous number of potential variants of glycosylated proteins (3). The most dominant plasma proteins, albumin and immunoglobulin (Ig)G, typically comprise up to 70% of the plasma proteome in abundance. To enable the majority of the remaining, far less abundant proteins to be better visualized by two-dimensional gel electrophoresis (2-DE), these two proteins must first be removed, or at least depleted in relative concentration. There are a number of currently available commercial products from a range of suppliers that enable albumin depletion by chemical affinity, exploiting the remarkable albumin-binding ability of structures closely related to the reactive dye molecule Cibacron blue 3GA (4), and the IgG binding properties of protein G (5). The blue dye has been shown to have a special affinity for proteins containing the dinucleotide fold, a structural feature that is common to several classes of proteins (4). Albumin can be separated from other plasma proteins using lectin affinity, as it is not normally glycosylated, while the majority of classical plasma proteins are. This approach allows both enrichment of lower-abundance proteins, and the study of differences in glycoprotein profiles (6).
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References
Steel, L. F., Trotter, M. G., Nakajima, P. B., Mattu, T. S., Gonye, G., and Block, T. (2003) Efficient and specific removal of albumin from human serum samples. Mol. Cell Proteomics 2, 262–270.
Tabar, L., Dean, P. B., Kaufman C. S., Duffy, S. W., and Chen, H. H. (2000) A new era in the diagnosis of breast cancer. Surg. Oncol. Clin. N. Am. 9, 233–277.
Anderson, N. L. and Anderson, N. G. (2002) The human plasma proteome, history character and diagnostic prospects. Mol. Cell Proteomics 1, 845–867.
Thompson, S. T., Cass, K. H., and Stellwagen, E. (1975) Blue dextran-sepharose: an affinity column for the dinucleotide fold. Proc. Nat. Acad. Sci. USA 72, 669–672.
Reis, K. J., Ayoub, E. M., and Boyle, M. D. P. (1984) Streptococcal Fc receptors. II. Comparison of the reactivity of a receptor from a group C streptococcus with staphylococcal protein A. J. Immunol. 132, 3098–3102.
Brzeski, H., Katenhusen, R. A., Sullivan, A. G., et al. (2003) Albumin depletion method for improved plasma glycoprotein analysis by 2-dimensional difference gel electrophoresis. Biotechniques 35, 1128–1132.
Steel, L. F., Trotter, M. G., Nakajima, P. B., Mattu, T. S., Gonye, G., and Block, T. (2003) Efficient and specific removal of albumin from human serum samples. Mol. Cell Proteomics 2, 262–270.
Pieper, R., Gatlin, C. L., Makusky A. J., et al. (2003) The human serum proteome: display of nearly 3700 chromatographically separated protein spots on two-dimensional electrophoresis gels and identification of 325 distinct proteins. Proteomics 3, 1345–1364.
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© 2005 Humana Press Inc., Totowa, NJ
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Sullivan, A.G., Russell, S., Brzeski, H., Somiari, R.I., Shriver, C.D. (2005). Serum or Plasma Sample Preparation for Two-Dimensional Gel Electrophoresis. In: Walker, J.M. (eds) The Proteomics Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-890-0:049
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DOI: https://doi.org/10.1385/1-59259-890-0:049
Publisher Name: Humana Press
Print ISBN: 978-1-58829-343-5
Online ISBN: 978-1-59259-890-8
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