Ultratrace Enrichment of Tyrosine Phosphorylated Peptides on an Imprinted Polymer

Abstract

Novel molecularly imprinted polymers (MIPs) designed to bind the side chain of phosphotyrosine can be used as artificial receptors for affinity-based enrichment of proteolytic peptides. In comparison with general enrichment methods for phosphorylated peptides such as TiO(2)-based methods, the pTyr-imprinted polymers offered high selectivity for pTyr-containing peptides down to the low fmol level. This suggests MIPs as a new tool for affinity-based proteomics.

Enrichment strategies for phosphoproteomics: state-of-the-art

“…Recently, phosphotyrosine MIP-based receptors have been developed using two urea-based monomers and an N,O-protected p-Tyr template. These receptors have been demonstrated to have high selectivity, sensitivity and affi nity for peptides containing the imprinted amino acid (pTyr) and hence could be used to Brought to you by | Western University Authenticated Download Date | 6/8/15 2:44 PM capture pTyr-containing peptides at the femtomole level in the presence of four orders of magnitude higher concentrations of other nonphosphorylated or pSer/Thr-phosphorylated peptide fragments (Helling et al 2011 ).…”

A “turn-on” fluorescent receptor for detecting tyrosine phosphopeptide using the surface imprinting procedure and the epitope approach

“…To analyze pTyr in molecular terms, versatile methods which can selectively recognized phosphorylated proteins and peptides have been established, such as pTyr specific antibodies (Lemeer et al, 2007), chemo-affinity enrichment based on metal mediated chelation (Muszynska et al, 1986), metal oxide affinity chromatography (Thingholm et al, 2006), electrostatic repulsion hydrophilic interaction chromatography (Alpert, 2007), and strong cation-exchange chromatography (Beausoleil et al, 2004). Although antibodies show high selectivity and specificity for pTyr recognition, they are costly and often affect the structure of proteins (Helling et al, 2011). Meanwhile, the protocols based on chemo-affinity are not specific for a given kind of phosphorylated residues, they generally simultaneously capture proteins or peptides containing phosphotyrosine, phosphoserine, phosphothreonine, or other amino acids including histidine and cysteine.…”

Automated Protein Biomarker Analysis: on-line extraction of clinical samples by Molecularly Imprinted Polymers

“…Precipitation polymerization is a very attractive synthetic method for the synthesis of MIP microspheres27282930313233. MIP microspheres of controlled size and porosity are obtained easily by the tuning of polymerization conditions343536 without the need for surfactants or stabilizers, delivering clean products with narrow particle size distributions.…”