Biology Reference
In-Depth Information
Chapter 13
2D DIGE Analysis of Serum After Fractionation
by ProteoMiner™ Beads
Cynthia Liang , Gek San Tan , and Maxey C. M. Chung
Abstract
Serum is a popular biofl uid 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 prefrac-
tionation methods have recently been developed with the view to reduce the dynamic range of these
proteins. These include both dye- and immunoaffi nity-based methods that are specifi cally 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. Introduction
Serum or plasma is routinely used for disease diagnosis and for
monitoring patient's health status. This is because blood circulates
the whole body and any changes in protein expression in serum or
plasma can refl ect the pathophysiology of the body. As the collec-
tion and processing of serum/plasma is also relatively noninvasive
and inexpensive, it is a popular biofl uid used for many protein
biomarker discovery projects. However, the downstream analysis
of serum/plasma is hampered severely by several high-abundant
proteins which often interfere with the separation and detection of
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