Summary
Human plasma is regarded the most complex and well-known clinical specimen that can be easily obtained; alterations in the levels of plasma proteins or their corresponding enzyme activities may reflect either a healthy or a diseased state. Given that there is no defined genomic information as to the intact protein components in plasma, protein profiling could be the first step toward its molecular characterization. Several problems exist in the analysis of plasma proteins, however. For example, the widest dynamic range of protein concentrations, the presence of high-abundance proteins, and post-translational modifications need to be considered before proteomic studies are undertaken. In particular, efficient depletion or pre-fractionation of high-abundance proteins is crucial for the identification of low-abundance proteins that may contain potential biomarkers. After the removal of high-abundance proteins, protein profiling can be initiated using two-dimensional electrophoresis (2DE), which has been widely used for displaying the differential proteome under specific physiological conditions. Here, we describe a typical 2DE procedure for plasma proteome under either a healthy or a diseased state (e.g., liver cancer) in which pre-fractionation and depletion are integral steps in the search for disease biomarkers.
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Abbreviations
- IEF:
-
Isoelectric Focusing
- IPG:
-
Immobilized pH Gradient
- TCA:
-
Trichloroacetic Acid
- FFE:
-
Free Flow Electrophoresis
- HPMC:
-
Hydroxypropyl Methylcellulose
- TBP:
-
Tributylphosphine
- 2DE:
-
2-dimensional Gel Electrophoresis
- BPB:
-
Bromophenol Blue
- CHCA:
-
α-cyano-4-hydroxycinnamic acid
- LTQ:
-
Linear Iontrap
- MALDI-TOF:
-
Matrix-assisted Laser Desorption Ionization - Time of Flight Mass Spectrometry
- HPPP:
-
Human Plasma Proteome Project.
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Acknowledgments
This study was supported by a grant from the Korean Health 21 R&D project, Ministry of Health & Welfare, Republic of Korea (A030003 to YKP).
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Cho, S.Y. et al. (2008). Protein Profiling of Human Plasma Samples by Two-Dimensional Electrophoresis. In: Vlahou, A. (eds) Clinical Proteomics. Methods in Molecular Biology™, vol 428. Humana Press. https://doi.org/10.1007/978-1-59745-117-8_4
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DOI: https://doi.org/10.1007/978-1-59745-117-8_4
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