The use of Cibacron Blue columns (HiTrapBlue) in proteome analysis for removal of plasma albumin, for facilitating biomarker discovery, has not borne any fruit. In fact, the visibility of low-abundance proteins was obscured. It is here reported that, upon albumin sequestering from plasma, there is adsorption, via hydrophobic interaction, of a substantial number of plasma proteins, which are lost for subsequent analysis if the blue resin is eluted via an ion shock (2. M NaCl) or with a somewhat more robust eluant (5. M urea, 2. M thiourea, 2% CHAPS, 2% sulphobetain 3-10) as recommended by manufacturers. Such treatments, in fact, release at most 25 to 30 unique gene products, including albumin. If, however, the Affigel-Blue resin, after elution with either of the two above eluants, is further eluted with boiling 4% SDS in 25. mM DTT, all the missing proteins (amounting to at least 112 unique species) are desorbed and biomarker analysis can be conducted in a correct way. It is also suggested that such blue-resin treatment could be coupled to ProteoMiner adsorption, this coupled treatment further enhancing the chances of success for discovery of low-abundance proteins.
Cibacron Blue and proteomics : the mystery of the platoon missing in action / F. Di Girolamo, P.G. Righetti, A. D'Amato, M.C.M. Chung. - In: JOURNAL OF PROTEOMICS. - ISSN 1874-3919. - 74:12(2011), pp. 2856-2865.
|Titolo:||Cibacron Blue and proteomics : the mystery of the platoon missing in action|
|Parole Chiave:||Biomarkers; Combinatorial peptide ligand libraries; Dye ligand affinity chromatography; Plasma; Biochemistry; Biophysics|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore CHIM/01 - Chimica Analitica
|Data di pubblicazione:||2011|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.jprot.2011.06.033|
|Appare nelle tipologie:||01 - Articolo su periodico|