Evaluation of a Solid-Supported Tagging Strategy for Mass Spectrometric Analysis of Peptides

Abstract

We have explored two divinylbenzene cross-linked polystyrene supports for use in a solid-supported N-terminal peptide tagging strategy. Resin-bound tags designed to be cleaved in a single step at the N-terminus of peptides have been devised and explored as peptide N-terminal tagging reagents (constructs) for subsequent mass spectrometric analysis. While the brominated tagging approach shows promise, the use of these specific solid supports has drawbacks, in terms of tagging reaction scale, for real applications in proteomics.

Fabrication and Application of Isotopically Labelled Gold Arrays for Multiplexed Peptide Analysis

“…Most solid‐supported approaches published to date involve a number of different steps: immobilisation of a label on the solid‐phase, capture of tryptic peptides by the labelling group, and then cleavage of the derivatised peptides from the solid‐phase for subsequent LC‐MS analysis . Routledge and co‐workers recently reported an alternative solid‐supported peptide labelling strategy that used peptidic N termini to cleave a solid‐phase‐immobilised brominated label, thus simplifying the workflow by avoiding the need for a separate release step . The use of a crosslinked polymer resin as the solid‐support resulted in an additional level of complexity in analysis due to the potential for nonspecific binding to the resin, but clearly demonstrated the benefit of bromine in the label, that it provides a stable mass spectrometry marker that allows labelled peptides to be readily distinguished from background signals.…”