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Serum Low-Molecular-Weight Protein Fractionation for Biomarker Discovery

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Molecular Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 823))

Abstract

Protein biomarkers provide the key diagnostic information for the detection of disease, risk of disease progression, and a patient’s likely response to drug therapy. Potential biomarkers exist in biofluids, such as serum, urine, and cerebrospinal fluid. Unfortunately, discovering and validating protein biomarkers are hindered by the presence of high-molecular-weight proteins, such as serum albumin and immunoglobulins, which comprise 90% of the proteins present in these samples. High-abundance, high-molecular-weight proteins mask the low-molecular-weight (LMW) proteins and peptides using conventional protein detection methods. Candidate biomarkers are believed to exist in very low concentrations and comprise less than 1% of serum proteins, and may be highly labile as well. Therefore, it is imperative to isolate and enrich LMW proteins from complex mixtures for biomarker discovery. This chapter describes a continuous ­elution electrophoresis method, based on molecular weight sieving, to isolate specific molecular weight fractions for mass spectrometric, western blotting, or protein array analysis.

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Author information

Authors and Affiliations

  1. Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA

    Amy J. VanMeter, Jonathan Hansen & Weidong Zhou

  2. Istituto Superiore di Sanità, Rome (RO), Italy

    Serena Camerini

  3. Istituto Superiore di Sanità-Ministero della Salute, Rome (RO), Italy

    Maria Letizia Polci

  4. Department of Experimental Medicine, University of L’Aquila, L’Aquila (AQ), Italy

    Alessandra Tessitore

  5. University of Virginia, Charlottesville, VA, USA

    Nishant Trivedi

  6. University of Virginia School of Medicine, Charlottesville, VA, USA

    Michael Heiby

  7. Children’s National Medical Center, Washington, DC, USA

    Yasmin Kamal

Authors
  1. Amy J. VanMeter
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  2. Serena Camerini
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  3. Maria Letizia Polci
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  4. Alessandra Tessitore
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  5. Nishant Trivedi
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  6. Michael Heiby
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  7. Yasmin Kamal
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  8. Jonathan Hansen
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  9. Weidong Zhou
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Corresponding author

Correspondence to Amy J. VanMeter .

Editor information

Editors and Affiliations

  1. Center for Applied Proteomics and, George Mason University, University Blvd. 10900, Manassas, 20110, Virginia, USA

    Virginia Espina

  2. Center for Applied Proteomics &, George Mason University, University Blvd. 10900, Manassas, 20110, Virginia, USA

    Lance A. Liotta

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© 2012 Springer Science+Business Media, LLC

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VanMeter, A.J. et al. (2012). Serum Low-Molecular-Weight Protein Fractionation for Biomarker Discovery. In: Espina, V., Liotta, L. (eds) Molecular Profiling. Methods in Molecular Biology, vol 823. Humana Press. https://doi.org/10.1007/978-1-60327-216-2_15

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  • DOI: https://doi.org/10.1007/978-1-60327-216-2_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-215-5

  • Online ISBN: 978-1-60327-216-2

  • eBook Packages: Springer Protocols

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