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Serum Low-Molecular-Weight Protein Fractionation for Biomarker Discovery

Protocol
First Online:
Part of the Methods in Molecular Biology book series (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.

Key words

Biomarker Continuous elution electrophoresis Fractionation Prep cell Low-molecular-weight protein Mass spectrometry Protein Serum 
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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Center for Applied Proteomics and Molecular MedicineGeorge Mason UniversityManassasUSA
  2. 2.Istituto Superiore di SanitàRome (RO)Italy
  3. 3.Istituto Superiore di Sanità-Ministero della SaluteRome (RO)Italy
  4. 4.Department of Experimental MedicineUniversity of L’AquilaL’Aquila (AQ)Italy
  5. 5.University of VirginiaCharlottesvilleUSA
  6. 6.University of Virginia School of MedicineCharlottesvilleUSA
  7. 7.Children’s National Medical CenterWashingtonUSA

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