Biology Reference
In-Depth Information
Chapter 12
Combination of Highly Effi cient Hexapeptide Ligand
Library-Based Sample Preparation with 2D DIGE for the
Analysis of the Hidden Human Serum/Plasma Proteome
Sonja Hartwig and Stefan Lehr
Abstract
Blood serum/plasma samples provide the major source to identify diagnostically relevant or treatment
response-related proteins. But its complexity and the enormous dynamic range hinders comprehensive
analysis. Therefore, more detailed analysis, in particular, of low-abundant peptides/proteins requires
extensive pre-fractionation, which frequently infl uences the native composition and may lead to a loss of
potentially important information. In order to overcome these barriers, we describe an innovative sample
preparation tool utilizing combinatorial hexapeptide ligand libraries to effi ciently enrich low-abundance
proteins with a simple protocol. In contrast to the most common approaches based on the immuno-
depletion of antibody-targeted high-abundance proteins, this technology concentrates low-abundance
proteins and concurrently reduces the high-abundance species. Thus, the dynamic range is compressed,
and low-abundance proteins become more easily detectable. We show how this sample preparation tech-
nique can be easily combined with 2D DIGE analysis to enable more comprehensive and quantitative
profi ling of complex biological samples.
Key words:
Serum profi ling, Sample pre-fractionation, Sample preparation, Low-abundant proteins,
Hexapeptide library beads, Combinatorial ligand libraries (CLL), Clinical proteomics, Biomarker
discovery
1. Introduction
Obesity, type 2 diabetes, and the cardio-metabolic syndrome are
epidemic diseases that represent a major threat to human health,
meaning a serious burden for our society. To understand the factors
causing such multifactorial diseases is one of the major driving
forces of biomedical research. In this research fi eld, quantitative
proteomic technology gives the opportunity to identify novel
biomarker enabling early detection, diagnosis, and monitoring of
disease progression.
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