Selective Enrichment of Phosphorylated Peptides by Monolithic Polymers Surface Imprinted with bis-Imidazolium Moieties by UV-Initiated Cryopolymerization

Abstract

Reversible protein phosphorylation on serine, threonine, and tyrosine residues is essential for fast, specific, and accurate signal transduction in cells. Up to now, the identification and quantification of phosphorylated amino acids, peptides and proteins continue to be one of the significant challenges in contemporary bioanalytical research. In this paper, a series of surface grafted monoliths in the capillary format targeting phosphorylated serine have been prepared by first synthesizing a monolithic core substrate material based on trimethylolpropane trimethacrylate, onto which a thin surface-imprinted layer was established by oriented photografting of a variety of mono-and bis-imidazolium host monomers at sub-zero temperature, using six different continuous or pulsed UV light sources. The imprinted monolith capillaries were evaluated in a capillary liquid chromatographic system connected to a mass spectrometer in order to test the specific retention of phosphorylated peptides. Site-specific recognition selectivity and specificity for phosphorylated serine was demonstrated when separating amino acids and peptides, proving that the optimized materials could be used as novel trapping media in affinity-based phosphoproteomic analysis.

Strategies of molecular imprinting-based solid-phase extraction prior to chromatographic analysis

“…However, their method performance, extremely depends on monolithic properties, so the optimization process of the porogen composition should be performed to arrive at similar porous properties. In the further work of this group, they developed the synthesis tactic to overcome the mentioned defect by polymerizing an appropriate monolithic substrate with well-defined porous properties and attaching groups on its pore surfaces, followed by grafting a thin MIP layer with the aid of these groups [90]. They prepared monoliths in the capillary format by trimethylolpropane trimethacrylate (TRIM) core on which MIP shell was decorated through oriented photografting of a variety of mono-and bis-imidazolium host monomers at subzero temperature, using six different continuous or pulsed UV light sources.…”

Epitope-imprinted polymers: Design principles of synthetic binding partners for natural biomacromolecules

“…This strategy has been successfully validated against the commonly used approaches based on TiO 2 and anti-pY antibodies (145) and has been used in combination with TiO 2 to selectively enrich a protein extracts from cancer cells in phosphopeptides before mass spectrometry analysis (146), suggesting that this method can be a valuable alternative for applications in phosphoproteomics research. This strategy has also been recently expanded to selectively bind phosphoserine (150,151) and phosphohistidine (152) residues, demonstrating its versatility.…”

High salt compatible oxyanion receptors by dual ion imprinting

“…Hence potent anion receptors can be prepared by polymerizing host monomers and a crosslinker in presence of the anion guest followed by guest removal. [11][12][13][14][15][16][17][18][19][20][21] The anion preorganizes the host which is covalently xed in a macromolecular scaffold with tunable local polarity. Permanent imprinted sites are achieved post template removal featuring enhanced affinity for the templated ion.…”