title: A novel heterotrifunctional peptide-based cross-linking reagent for facile access to bioconjugates. Applications to peptide fluorescent labelling and immobilisation.
abstract: A convenient, versatile and straightforward synthesis of a novel heterotrifunctional peptide-based linker molecule is described. This generic bio-labelling reagent contains an amine-reactive N-hydroxysuccinimidyl carbamate moiety, an aldehyde/ketone-reactive aminooxy group and a thiol group with a propensity to form urea, oxime and thioether linkages respectively. The full chemical orthogonality between the free aminooxy and thiol functionalities was demonstrated through the preparation of a fluorescent reagent suitable for the selective staining of a carboxaldehyde-modified surface by means of oxime ligation. The absence of reactivity of these two functions toward the nucleophile-sensitive active carbamate was obtained by using temporary aminooxy- and thiol-protecting groups removable under mild conditions. The utility of the linker molecule to cross-link three different molecular partners has been illustrated by the preparation of fluorescent tripod-functionalised surfaces which may be useful in developing new peptide microarrays and related immunosensors.
authors.affiliation: Equipe de Chimie Bio-Organique, COBRA - CNRS UMR 6014, rue Lucien Tesnière, 76131, Mont-Saint-Aignan, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Clavé
authors.forename: Hervé
authors.initials: H
authors.lastname: Boutal
authors.forename: Antoine
authors.initials: A
authors.lastname: Hoang
authors.forename: François
authors.initials: F
authors.lastname: Perraut
authors.forename: Hervé
authors.initials: H
authors.lastname: Volland
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Renard
authors.forename: Anthony
authors.initials: A
authors.lastname: Romieu
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D015200:Fluoroimmunoassay / Q000379:methods*; D013373:Substance P / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D013438:Sulfhydryl Compounds; D013373:Substance P