Sample preparation techniques for the untargeted LC-MS-based discovery of peptides in complex biological matrices.
Although big progress has been made in sample pretreatment over the last years, there are still considerable limitations when it comes to overcoming complexity and dynamic range problems associated with peptide analyses from biological matrices. Being the little brother of proteomics, peptidomics is a relatively new field of research aiming at the direct analysis of the small proteins, called peptides, many of which are not amenable for typical trypsin-based analytics. In this paper, we present an overview of different techniques and methods currently used for reducing a sample's complexity and for concentrating low abundant compounds to enable successful peptidome analysis. We focus on techniques which can be employed prior to liquid chromatography coupled to mass spectrometry for peptide detection and identification and indicate their advantages as well as their shortcomings when it comes to the untargeted analysis of native peptides from complex biological matrices.
Laboratory for Analytical Biotechnology and Innovative Peptide Biology, Department of Biotechnology, Delft University of Technology, Julianalaan 67, 3706 AV Zeist, The Netherlands. i.m.o.finoulst@tudelft.nl
Inez
I
Finoulst
Martijn
M
Pinkse
William
W
Van Dongen
Peter
P
Verhaert
D053000:Analytic Sample Preparation Methods / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D045424:Complex Mixtures / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis*; D040901:Proteomics / Q000379:methods
D045424:Complex Mixtures; D010455:Peptides