Highly selective enrichment of phosphopeptides by titanium (IV) attached monodisperse-porous poly(vinylphosphonic acid-co-ethylene dimethacrylate) microspheres


SALIMI K., Usta D. D. , ÇELİKBIÇAK Ö., PINAR A., SALİH B., Tuncel A.

JOURNAL OF CHROMATOGRAPHY A, vol.1496, pp.9-19, 2017 (Peer-Reviewed Journal) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 1496
  • Publication Date: 2017
  • Doi Number: 10.1016/j.chroma.2017.03.052
  • Journal Name: JOURNAL OF CHROMATOGRAPHY A
  • Journal Indexes: Science Citation Index Expanded, Scopus
  • Page Numbers: pp.9-19
  • Keywords: Vinylphosphonic acid, Monodisperse-porous microspheres, Phosphopeptide enrichment, Human serum, Bovine serum albumin, ION AFFINITY-CHROMATOGRAPHY, METAL-ORGANIC FRAMEWORK, MODIFIED SILICA NANOPARTICLES, IONIZATION MASS-SPECTROMETRY, PHOSPHOPROTEOME ANALYSIS, MAGNETIC NANOPARTICLES, SENSITIVE DETECTION, BIOLOGICAL SAMPLES, SIGNALING NETWORKS, CAPTURE

Abstract

A seeded polymerization protocol was developed for the synthesis of monodisperse-porous poly(vinylphosphonic acid-co-ethylene dimethacrylate), [poly(VPA-co-EDMA)] microspheres with superior porous properties. The protocol allowed the direct synthesis of phosphonic acid functionalized porous microspheres with the mean size of similar to 4 mu m and the specific surface area of 420 m(2) g(-1) without applying any complicated post-derivatization protocol for the attachment of phosphonic acid group. The phosphonic acid content of poly(VPA-co-EDMA) microspheres was determined as 1.5 mmol H2PO3 g(-1) microspheres. Ti(IV) ions were attached onto the microspheres via metal-chelate complex formation by phosphonate-groups and Ti(IV) carrying monodisperse-porous poly(vinylphosphonic acid-co-ethylene dimethacrylate), [Ti(IV)@poly(VPA-co-EDMA)]microspheres were obtained as a new sorbent for phosphopeptide enrichment via immobilized metal affinity chromatography. The phosphopeptides in the enriched samples were identified by matrix-assited laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Four different phosphopeptides were detected with extremely high intensity by the treatment of beta-casein digest prepared with the concentration of 10 fmol/mL with Ti(IV)@poly(VPA-co-EDMA) microspheres. Highly selective enrichment of phosphopeptides was also successfully carried out even at trace amounts in a complex mixture of digested proteins (molar ratio of (beta-casein to BSA, 1:1000) and eight different phosphorylated peptides from BSA digest were successfully identified. Moreover, four highly intense signals of the phosphopeptides in human serum were observed with high S/N ratio and clear background after enrichment by using Ti(IV) poly(VPA-co-EDMA) microspheres. (C) 2017 Elsevier B.V. All rights reserved.