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Molecularly imprinted polymers synthesized via template immobilization on fumed silica nanoparticles for the enrichment of phosphopeptides

Nogueira Duarte, Mariana LU ; Subedi, Prabal; Yilmaz, Ecevit LU ; Marcus, Katrin; Laurell, Thomas LU and Ekström, Simon LU (2018) In Journal of Molecular Recognition 31(3). p.2677-2677
Abstract

Phosphorylation is a protein post-translational modification (PTM) that plays an important role in cell signaling, cell differentiation, and metabolism. The hyper phosphorylated forms of certain proteins have been appointed as biomarkers for neurodegenerative diseases, and phosphorylation-related mutations are important for detecting cancer pathways. Due to the low abundance of phosphorylated proteins in biological fluids, sample enrichment is beneficial prior to detection. Thus, a need to find new strategies for enriching phosphopeptides has emerged. Molecularly imprinted polymers (MIPs) are synthetic polymeric materials manufactured to exhibit affinity for a target molecule. In this study, MIPs have been synthesized using a new... (More)

Phosphorylation is a protein post-translational modification (PTM) that plays an important role in cell signaling, cell differentiation, and metabolism. The hyper phosphorylated forms of certain proteins have been appointed as biomarkers for neurodegenerative diseases, and phosphorylation-related mutations are important for detecting cancer pathways. Due to the low abundance of phosphorylated proteins in biological fluids, sample enrichment is beneficial prior to detection. Thus, a need to find new strategies for enriching phosphopeptides has emerged. Molecularly imprinted polymers (MIPs) are synthetic polymeric materials manufactured to exhibit affinity for a target molecule. In this study, MIPs have been synthesized using a new approach based on the use of fumed silica as sacrificial support acting as solid porogen with the template (phosphotyrosine) immobilized on its surface. Phosphotyrosine MIPs were tested against a mixture of peptides and phosphopeptides by performing micro-solid phase extraction using MIPs (μMISPE) packed in a pipette tip. First, the capability of the materials to preferentially enrich phosphopeptides was evaluated. In a next step, the enrichment of phosphopeptides from a whole-cell lysate of human embryonic kidney (HEK) 293T cells was performed. The eluates were analyzed using MALDI-MS in the first case and with nano-HPLC-ESI-MS/MS in the second case. The results showed that the MIPs provided affinity for phosphopeptides, binding preferentially to multi-site phosphorylated peptides. The MIPs could enrich phosphopeptides in over 10-fold compared with the number of phosphopeptides found in a cell lysate without enrichment.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Fumed silica nanoparticles, Molecularly imprinted polymers, Phosphopeptide enrichment, Pore-size distribution, Surface imprinting
in
Journal of Molecular Recognition
volume
31
issue
3
pages
2677 - 2677
publisher
John Wiley & Sons
external identifiers
  • scopus:85031107645
ISSN
0952-3499
DOI
10.1002/jmr.2677
language
English
LU publication?
yes
id
01eef804-cb4f-4ae5-90ad-23cc2a545096
date added to LUP
2017-10-30 08:43:31
date last changed
2018-03-01 04:30:00
@article{01eef804-cb4f-4ae5-90ad-23cc2a545096,
  abstract     = {<p>Phosphorylation is a protein post-translational modification (PTM) that plays an important role in cell signaling, cell differentiation, and metabolism. The hyper phosphorylated forms of certain proteins have been appointed as biomarkers for neurodegenerative diseases, and phosphorylation-related mutations are important for detecting cancer pathways. Due to the low abundance of phosphorylated proteins in biological fluids, sample enrichment is beneficial prior to detection. Thus, a need to find new strategies for enriching phosphopeptides has emerged. Molecularly imprinted polymers (MIPs) are synthetic polymeric materials manufactured to exhibit affinity for a target molecule. In this study, MIPs have been synthesized using a new approach based on the use of fumed silica as sacrificial support acting as solid porogen with the template (phosphotyrosine) immobilized on its surface. Phosphotyrosine MIPs were tested against a mixture of peptides and phosphopeptides by performing micro-solid phase extraction using MIPs (μMISPE) packed in a pipette tip. First, the capability of the materials to preferentially enrich phosphopeptides was evaluated. In a next step, the enrichment of phosphopeptides from a whole-cell lysate of human embryonic kidney (HEK) 293T cells was performed. The eluates were analyzed using MALDI-MS in the first case and with nano-HPLC-ESI-MS/MS in the second case. The results showed that the MIPs provided affinity for phosphopeptides, binding preferentially to multi-site phosphorylated peptides. The MIPs could enrich phosphopeptides in over 10-fold compared with the number of phosphopeptides found in a cell lysate without enrichment.</p>},
  author       = {Nogueira Duarte, Mariana and Subedi, Prabal and Yilmaz, Ecevit and Marcus, Katrin and Laurell, Thomas and Ekström, Simon},
  issn         = {0952-3499},
  keyword      = {Fumed silica nanoparticles,Molecularly imprinted polymers,Phosphopeptide enrichment,Pore-size distribution,Surface imprinting},
  language     = {eng},
  number       = {3},
  pages        = {2677--2677},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Molecular Recognition},
  title        = {Molecularly imprinted polymers synthesized via template immobilization on fumed silica nanoparticles for the enrichment of phosphopeptides},
  url          = {http://dx.doi.org/10.1002/jmr.2677},
  volume       = {31},
  year         = {2018},
}