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Poly(ethylene glycol) functionalization of monolithic poly(divinyl benzene) for improved miniaturized solid phase extraction of protein-rich samples
journal contribution
posted on 2017-03-01, 00:00 authored by Esme Candish, Aminreza Khodabandeh, Marianne Gaborieau, Thomas Rodemann, Robert ShellieRobert Shellie, Andrew A Gooley, Emily F HilderNon-specific protein adsorption on hydrophobic solid phase extraction (SPE) adsorbents can reduce the efficacy of purification. To improve sample clean-up, poly(divinyl benzene) (PDVB) monoliths grafted with hydrophilic polyethylene glycol methacrylate (PEGMA) were developed. Residual vinyl groups (RVGs) of the PDVB were employed as anchor points for PEGMA grafting. Two PEGMA monomers, M n 360 and 950, were compared for graft solutions containing 5-20% monomer. Protein binding was qualitatively screened using fluorescently labeled human serum albumin (HSA) to determine optimal PEGMA concentration. The fluorescent signal of PDVB was reduced for PDVB-g-PEGMA360 (10%) and PDVB-g-PEGMA950 (20%). The PEGMA content (w/w%) was quantified by solid state 1H NMR to be 29.9 ± 1.6% for PDVB-g-PEGMA360 and 7.7 ± 1.2% for PDVB-g-PEGMA950. To assess adsorbent performance breakthrough curves for PDVB, PDVB-g-PEGMA360 and PDVB-g-PEGMA950 were compared. The breakthrough volume (V B) and shape of the curve for PDVB-g-PEGMA950 were maintained relative to PDVB (2.3 and 2.8 mL, respectively). A reduced V B of 0.5 mL and shallow breakthrough curve indicated PDVB-g-PEGMA360 was not suitable for SPE. A high ibuprofen recovery of 92 ± 0.30 and 78 ± 0.93% was seen for PDVB and PDVB-g-PEGMA950, respectively. Protein adsorption was reduced from 31 ± 2.41 to 12 ± 0.49% for PDVB and PDVB-g-PEGMA950, respectively. SPE of ibuprofen from plasma was compared for PDVB and PDVB-g-PEGMA950 by at-line electrospray ionization mass spectrometry (ESI-MS). PDVB-g-PEGMA950 demonstrated a threefold increase in assay sensitivity indicating a superior analyte purification.
History
Journal
Analytical and bioanalytical chemistryVolume
409Issue
8Pagination
2189 - 2199Publisher
Springer VerlagLocation
Berlin, GermanyPublisher DOI
ISSN
1618-2642eISSN
1618-2650Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2017, Springer-Verlag Berlin HeidelbergUsage metrics
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BiocompatibleGraftingPorous polymer monolithSample preparationSolid phase extractionFluorescent DyesMicroscopy, Electron, ScanningMiniaturizationNuclear Magnetic Resonance, BiomolecularPolyethylene GlycolsPolymersProteinsVinyl CompoundsScience & TechnologyLife Sciences & BiomedicinePhysical SciencesBiochemical Research MethodsChemistry, AnalyticalBiochemistry & Molecular BiologyChemistryPERFORMANCE LIQUID-CHROMATOGRAPHYPOROUS POLYMER MONOLITHSBIOLOGICAL-FLUIDSSTATIONARY PHASESDIRECT-INJECTIONSILICA SUPPORTSSTATE NMRMEDIAMETHACRYLATESEPARATION
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