Hierarchically templated beads with tailored pore structure for phosphopeptide capture and phosphoproteomics. Issue 28 (17th March 2017)

• Record Type: Journal Article
• Authors: Wierzbicka, Celina; Torsetnes, Silje B.; Jensen, Ole N.; Shinde, Sudhirkumar; Sellergren, Börje
• Is Part Of: RSC advances. Volume 7:Issue 28(2017)
• Page Start: 17154
• Page End: 17163
• Publication Date: 2017-03-17
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c7ra00385d
• Languages: English
• ISSNs: 2046-2069

Abstract

Abstract : Phosphotyrosine selective beads prepared by polymer templating at two length scales results in improved capture of larger sized peptide fragments from tryptic protein digests. Abstract : Two templating approaches to produce imprinted phosphotyrosine capture beads with a controllable pore structure are reported and compared with respect to their ability to enrich phosphopeptides from a tryptic peptide mixture. The beads were prepared by the polymerization of urea-based host monomers and crosslinkers inside the pores of macroporous silica beads with both free and immobilized template. In the final step the silica was removed by fluoride etching resulting in mesoporous polymer replicas with narrow pore size distributions, pore diameters ≈ 10 nm and surface area > 260 m 2 g −1 . The beads displayed pronounced phosphotyrosine affinity and selectivity in binding tests using model peptides in acetonitrile rich solutions with a performance surpassing solution polymerized bulk imprinted materials. Tests of the beads for the enrichment of phosphopeptides from tryptic digests of twelve proteins revealed both pY/pS and pY/Y selectivity. This was reflected in a nearly 6-fold increase in the enrichment factor of a 23- mer pY-peptide and pY/pS normalized intensity ratios up to 1.5, when comparing the template mesoporous beads with the bulk materials.