The proteome buccaneers: how to unearth your treasure chest via combinatorial peptide ligand libraries.
The latest advances in combinatorial peptide ligand libraries, with their unique performance in discovering low-abundance species in proteomes, are reviewed here. Explanations of mechanism, potential applications, capture of proteomes at different pH values to enhance the total catch and quantitative elutions, such as boiling in the presence of 5% sodium dodecyl sulfate and 3% dithiothreitol are included. The reproducibility of protein capture among different experiments with the same batch of beads or with different batches is also reported to be very high, with coefficient of variations in the order of 10-20%. Miniaturized operations, consisting of capture with as little as 20 or even 5 microl of peptide beads are reported, thus demonstrating that the described technology could be exploited for routine biomarker discovery in a biomedical environment. Finally, it is shown that the signal of captured proteins is linear over approximately three orders of magnitude, ranging from nM to microM, thus ensuring that differential quantitative proteomics for biomarker discovery can be fully implemented, providing species do not saturate their ligands.
Righetti
Pier Giorgio
PG
Department of Chemistry, Materials and Chemical Engineering Giulio Natta, Via Mancinelli 7, Politecnico di Milano, Milano, Italy. piergiorgio.righetti@polimi.it
Boschetti
Egisto
E
Kravchuk
Alexander V
AV
Fasoli
Elisa
E
D015415:Biomarkers / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D006801:Humans; D008024:Ligands; D019151:Peptide Library; D020543:Proteome / Q000378:metabolism*
D015415:Biomarkers; D008024:Ligands; D019151:Peptide Library; D020543:Proteome