Abstract
The present report tries to assess the possibility of performing capture of proteomes via combinatorial peptide ligand libraries (CPLL) in a two-dimensional (2D) mode, i.e. via orthogonal complementarity in the capture phase. To that aim, serum proteins are captured at physiological pH either at low ionic strength (25. mM NaCl) or at high concentrations of lyotropic salts of the Hofmeister series (1. M ammonium sulphate) favouring hydrophobic interaction. Indeed such 2D mechanisms seems to be operative, since 52% of the captured proteins are common to the two capture modes, 20% are specific only of the "ionic" interaction mode and 28% are found only in the "hydrophobically" driven interaction. As an additional bonus, losses of protein species from the initial sample, one of the major drawbacks of CPLLs, are diminished to about 5% and are found only in the ionic capture, whereas the hydrophobically engendered capture is loss-free.
| Original language | English |
|---|---|
| Pages (from-to) | 106-112 |
| Number of pages | 7 |
| Journal | Journal of Chromatography A |
| Volume | 1297 |
| DOIs | |
| Publication status | Published - Jul 5 2013 |
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Keywords
- Combinatorial peptide ligand libraries
- Human serum
- Hydrophobic capture
- Ionic capture
- Mass spectrometry
ASJC Scopus subject areas
- Analytical Chemistry
- Organic Chemistry
- Biochemistry
Cite this
Combinatorial ligand libraries as a two-dimensional method for proteome analysis. / Santucci, Laura; Candiano, Giovanni; Petretto, Andrea; Lavarello, Chiara; Bruschi, Maurizio; Ghiggeri, Gian Marco; Citterio, Attilio; Righetti, Pier Giorgio.
In: Journal of Chromatography A, Vol. 1297, 05.07.2013, p. 106-112.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Combinatorial ligand libraries as a two-dimensional method for proteome analysis
AU - Santucci, Laura
AU - Candiano, Giovanni
AU - Petretto, Andrea
AU - Lavarello, Chiara
AU - Bruschi, Maurizio
AU - Ghiggeri, Gian Marco
AU - Citterio, Attilio
AU - Righetti, Pier Giorgio
PY - 2013/7/5
Y1 - 2013/7/5
N2 - The present report tries to assess the possibility of performing capture of proteomes via combinatorial peptide ligand libraries (CPLL) in a two-dimensional (2D) mode, i.e. via orthogonal complementarity in the capture phase. To that aim, serum proteins are captured at physiological pH either at low ionic strength (25. mM NaCl) or at high concentrations of lyotropic salts of the Hofmeister series (1. M ammonium sulphate) favouring hydrophobic interaction. Indeed such 2D mechanisms seems to be operative, since 52% of the captured proteins are common to the two capture modes, 20% are specific only of the "ionic" interaction mode and 28% are found only in the "hydrophobically" driven interaction. As an additional bonus, losses of protein species from the initial sample, one of the major drawbacks of CPLLs, are diminished to about 5% and are found only in the ionic capture, whereas the hydrophobically engendered capture is loss-free.
AB - The present report tries to assess the possibility of performing capture of proteomes via combinatorial peptide ligand libraries (CPLL) in a two-dimensional (2D) mode, i.e. via orthogonal complementarity in the capture phase. To that aim, serum proteins are captured at physiological pH either at low ionic strength (25. mM NaCl) or at high concentrations of lyotropic salts of the Hofmeister series (1. M ammonium sulphate) favouring hydrophobic interaction. Indeed such 2D mechanisms seems to be operative, since 52% of the captured proteins are common to the two capture modes, 20% are specific only of the "ionic" interaction mode and 28% are found only in the "hydrophobically" driven interaction. As an additional bonus, losses of protein species from the initial sample, one of the major drawbacks of CPLLs, are diminished to about 5% and are found only in the ionic capture, whereas the hydrophobically engendered capture is loss-free.
KW - Combinatorial peptide ligand libraries
KW - Human serum
KW - Hydrophobic capture
KW - Ionic capture
KW - Mass spectrometry
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U2 - 10.1016/j.chroma.2013.04.065
DO - 10.1016/j.chroma.2013.04.065
M3 - Article
C2 - 23726082
AN - SCOPUS:84878951465
VL - 1297
SP - 106
EP - 112
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
ER -