2D DIGE Analysis of Serum After Fractionation by ProteoMiner™ Beads

  • Cynthia Liang
  • Gek San Tan
  • Maxey C. M. ChungEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 854)


Serum is a popular biofluid used for many protein biomarker discovery projects since the collection and processing of serum/plasma is relatively noninvasive and inexpensive. Unfortunately, the downstream analysis of serum/plasma is hampered severely by several high-abundant proteins which often interfere with the separation and detection of many of the proteins of lower abundance. Thus, a number of prefractionation methods have recently been developed with the view to reduce the dynamic range of these proteins. These include both dye- and immunoaffinity-based methods that are specifically designed to remove serum albumin. In this chapter, we describe an alternative method using ProteoMiner™ or Equalizer beads that is aimed at overcoming this problem in serum. This method uses a combinatorial library of hexapeptides bound to beads and works by binding proteins until saturation is reached. Thus, the high-abundant proteins will reach saturation quickly, while the lower-abundant proteins continue to bind. This results in a dramatic depletion of the most abundant proteins, with a concurrent concentration of the middle- to low-abundant proteins.

Key words

Serum Equalizer beads ProteoMiner™ 2D DIGE 


  1. 1.
    States DJ, Omenn GS, Blackwell TW et al (2006) Challenges in deriving high-confidence protein identifications from data gathered by a HUPO plasma proteome collaborative study. Nat. Biotechnol. 24:333–338.PubMedCrossRefGoogle Scholar
  2. 2.
    Lowenthal MS, Mehta AI, Frogale et al (2005) Analysis of albumin-associated peptides and proteins from ovarian cancer patients. Clin. Chem. 51:1933–1945.Google Scholar
  3. 3.
    Zhou M, Lucas DA, Chan KC et al (2004) An investigation into the human serum “interactome”. Electrophoresis 25:1289–1298.PubMedCrossRefGoogle Scholar
  4. 4.
    Gundry R, Fu Q, Jelinek C (2007) Investigation of an albumin-enriched fraction of human serum and its albuminome. Proteomics Clin. Appl. 1:73–88.PubMedCrossRefGoogle Scholar
  5. 5.
    Mehta AI, Ross S, Lowenthal MS, Fusaro V et al (2003) Biomarker amplification by serum carrier protein binding. Dis. Markers 19:1–10.PubMedGoogle Scholar
  6. 6.
    Gundry RL, Cotter RJ (2007) From Diagnosis to Therapy. In: Dunn, MJ, Van Eyk, JE (Eds.) Clinical Proteomic, Wiley, London p. 263.Google Scholar
  7. 7.
    Gundry RL, Fu Q, Jelinek CA et al (2007) Investigation of an albumin-enriched fraction of human serum and its albuminome. Proteomics Clin. Appl. 1:73–88.PubMedCrossRefGoogle Scholar
  8. 8.
    Thulasiraman V, Lin S, Gheorghiu L et al (2005) Reduction of the concentration difference of proteins in biological liquids using a library of combinatorial ligands. Electrophoresis 26: 3561–3571.PubMedCrossRefGoogle Scholar
  9. 9.
    Marrocco C, Rinalducci S, Mohamadkhan A et al (2010) Plasma gelsolin protein: a candidate biomarker for hepatitis B-associated liver cirrhosis identified by proteomic approach. Blood Transfus 8: Suppl 3:s105–112.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Cynthia Liang
    • 1
  • Gek San Tan
    • 1
  • Maxey C. M. Chung
    • 2
    Email author
  1. 1.Department of Biological Science, Faculty of ScienceNational University of SingaporeSingaporeSingapore
  2. 2.Department of Biochemistry and Biological Sciences, Yong Loo Lin School of Medicine and Faculty of ScienceNational University of SingaporeSingaporeSingapore

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