title: Combinatorial peptide libraries: robotic synthesis and analysis by nuclear magnetic resonance, mass spectrometry, tandem mass spectrometry, and high-performance capillary electrophoresis techniques.
abstract: Combinatorial peptide libraries are a new source of compounds from which a large number of pharmacological leads will emerge in the next few years. A large body of literature shows that this approach is of considerable interest in many areas of biological sciences for the search of enzyme substrates and inhibitors, of receptor agonists or antagonists, or of antigen sequences. Nevertheless, the analytical investigation of such complex mixtures as libraries which contain up to millions of individual molecules is still poorly documented. In this work, we present analytical solutions for their characterization. NMR and tandem mass spectrometry (MS/MS) can provide an in deep description of any type of combinatorial libraries, while MS and high-performance capillary electrophoresis can bring a rapid and overall information at the routine level, sufficient to ensure a first assessment of their composition. MS in the fast atom bombardment mode was used to describe the libraries O1X2X3X4X5 or O1X2X3X4 (Oi and Xi are fixed and random residue in position i, respectively). Advantage was taken of the high proton affinity of arginine and of its induction of charge remote fragmentation to interpret the MS spectra of whole libraries and neutral losses (MS/MS) in the model sublibraries ArgGlyX3X4 and NipValX3X4 (Nip,4-nitrophenylalanine). Two-dimensional NMR allowed the incorporation of the individual residues during synthesis to be tested in 24 sublibraries O1X2X3X4. While from the pharmacological point of view, impressive discoveries made with combinatorial peptide libraries have already been reported, our results show that they should be complemented by appropriate analytical tools, crucial for the proper characterization and exploitation of these libraries.
authors.affiliation: Division des Peptides, Institut de Recherches Servier, Suresnes, France.
authors.forename: J A
authors.initials: JA
authors.lastname: Boutin
authors.forename: P
authors.initials: P
authors.lastname: Hennig
authors.forename: P H
authors.initials: PH
authors.lastname: Lambert
authors.forename: S
authors.initials: S
authors.lastname: Bertin
authors.forename: L
authors.initials: L
authors.lastname: Petit
authors.forename: J P
authors.initials: JP
authors.lastname: Mahieu
authors.forename: B
authors.initials: B
authors.lastname: Serkiz
authors.forename: J P
authors.initials: JP
authors.lastname: Volland
authors.forename: J L
authors.initials: JL
authors.lastname: Fauchère
mesh_terms: D000595:Amino Acid Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D015723:Gene Library; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics*; D012371:Robotics
chemical_list: D010455:Peptides