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Proteomics. 2010 Apr;10(7):1416-25. doi: 10.1002/pmic.200900431.

Comparison of protein enrichment strategies for proteome analysis of plasma.

Author information

1
Pfizer Global Research and Development, Pfizer Inc, Ann Arbor, MI, USA. julia.bandow@rub.de

Abstract

Efforts to discover protein biomarkers in plasma are hampered by the high abundance of few proteins, which interfere with the detection of low-abundant proteins. Different commercially available protein-partitioning products were tested for their ability to lower the detection limit of proteins in 2-D gels. Immuno-depletion using polyclonal antibodies raised against the proteins of highest abundance (Seppro IgY14 System) was compared with a two-step immuno-depletion strategy, where depletion with the Seppro IgY14 column was followed by depletion with the Seppro IgY-SuperMix system. The third strategy tested was protein pre-fractionation using the ProteoMiner kit, where proteins compete for binding sites on bead-bound peptide hexamers with different binding properties. The pre-fractionated protein samples were analyzed using 2-DE, which revealed stunning differences in protein patterns. However, detectable protein spots in the different plasma fractions contained exclusively high-abundant proteins normally present in plasma at concentrations between 1 microg and 40 mg/mL.

PMID:
20127685
DOI:
10.1002/pmic.200900431
[Indexed for MEDLINE]

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