Free flow electrophoresis (FFE) is a fractionation method, based on isoelectric focusing (IEF). We validate the reproducibility of the method and show that it can be applied by biobanks in order to fractionate fluid biospecimens efficiently and reproducibly and to facilitate downstream proteomic applications. We also propose a simple method allowing researchers to assess the reproducibility of each FFE run.
Get full access to this article
View all access options for this article.
References
1.
AndersonNL, PolanskiM, PieperR, GatlinT, TirumalaiRS, ConradsTP, VeenstraTD, AdkinsJN, PoundsJG, FaganR, LobleyA. The human plasma proteome: A nonredundant list developed by combination of four separate sources. Mol Cell Proteomics, 2004; 3:311–326.
2.
HannigK. Die trägerfreie kontinuierliche Elektrophorese und ihre Anwendung. Fresenius Zeitscr Anal Chem, 1961; 181:244–274.
HoffmannP, JiH, MoritzRL, ConnollyLM, FrecklingtonDF, LaytonMJ, EddesJS, SimpsonRJ. Continuous free flow electrophoresis separation of cytosolic proteins from the human colon carcinoma cell line LIM 1215: A non two-dimensional gel electrophoresis-based proteome analysis strategy. Proteomics, 2001; 1:807–818.
5.
ChoSY, LeeEY, LeeJS, KimHY, ParkJM, KwonMS, ParkYK, LeeHJ, KangMJ, KimJY, YooJS, ParkDJ, ChoJW, KimHS, PaikYK. Efficient prefractionation of low-abundance proteins in human plasma and construction of a two-dimensional map. Proteomics, 2005; 5:3386–3396.
6.
MoritzRL, ClippingdaleAB, KappEA, EddesJS, JiH, GilbertS, ConnollyLM, SimpsonRJ. Application of 2-D free-flow electrophoresis/RP-HPLC for proteomic analysis of human plasma depleted of multi high-abundance proteins. Proteomics, 2005; 5:3402–3414.
7.
MoritzRL, JiH, SchützF, ConnollyLM, KappEA, SpeedTP, SimpsonRJ. A proteome strategy for fractionating proteins and peptides using continuous free-flow electrophoresis coupled off-line to reversed-phase high-performance liquid chromatography. Anal Chem, 2004; 76:4811–4824.
WildgruberR, YiJ, NissumM, EckerskornC, WeberG. Free-Flow Electrophoresis system for plasma proteomic applications. Methods Mol Biol, 2008; 424:287–300.
11.
BrouilletteCG, AnantharamaiahGM, EnglerJA, BorhaniDW. Structural models of human apolipoprotein A-I: A critical analysis and review. Biochim Biophys Acta, 2001; 1531:4–46.
12.
AjeesAA, AnantharamaiahGM, MishraVK, HussainMM, MurthyHM. Crystal structure of human apolipoprotein A-I: Insights into its protective effect against cardiovascular diseases. Proc Natl Acad Sci USA, 2006; 103:2126–2131.
13.
SegelmarkM, PerssonB, HellmarkT, WieslanderJ. Binding and inhibition of myeloperoxidase (MPO): A major function of ceruloplasmin?Clin Exp Immunol, 1997; 108:167–174.
14.
KangJH. Modification and inactivation of human ceruloplasmin by oxidized DOPA. Bull Korean Chem Soc, 2004; 25:625–628.
15.
KalshekerN. Alpha-antitrypsin: Structure, function and molecular biology of the gene. Biosci Rep, 1989; 9:2129–2138.