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On-chip solid phase extraction and enzyme digestion using cationic PolyE-323 coatings and porous polymer monoliths coupled to electrospray mass spectrometry

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1016/j.chroma.2011.04.027
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Affiliation
  1. National Research Council of Canada. National Institute for Nanotechnology
FormatText, Article
SubjectCapillary wall; Cationic charges; Cytochrome C; Electrokinetic; Electroosmotic flow; Electrospray mass spectrometry; Electrosprays; Enzyme digestion; Glass chips; High quality; Ionization mass spectrometry; Linear calibration curve; Migration time; Model proteins; Monolithic column; Monolithic columns; Monolithic polymers; On chips; Peptide adsorption; Polymer monoliths; Porous polymer monoliths; Porous polymers; Pre-concentration; Protein adsorption; Solid phase extraction; Solid-phase extraction columns; Solution phase; Stop flow; Surface coatings; Trypsin digestion; Adsorption; Chlorine compounds; Electroosmosis; Electrophoresis; Electrospray ionization; Ion chromatography; Mass spectrometry; Microfluidics; Monolithic integrated circuits; Peptides; Plastic coatings; Polymers; Extraction; cation; methacrylic acid; PolyE 323; polymer; unclassified drug; adsorption; calibration; capillary electrophoresis; electroosmosis; electrospray mass spectrometry; enzyme degradation; hydrophobicity; material coating; microfluidic analysis; microfluidics; polymerization; porosity; separation technique; solid phase extraction; Electroosmosis; Microfluidic Analytical Techniques; Microscopy, Electron, Scanning; Peptide Fragments; Polymerization; Polymethacrylic Acids; Proteins; Solid Phase Extraction; Spectrometry, Mass, Electrospray Ionization; Trypsin
Abstract
We evaluate the compatibility and performance of polymer monolith solid phase extraction beds that incorporate cationic charge, with a polycationic surface coating, PolyE-323, fabricated within microfluidic glass chips. The PolyE-323 is used to reduce protein and peptide adsorption on capillary walls during electrophoresis, and to create anodal flow for electrokinetically driven nano-electrospray ionization mass spectrometry. A hydrophobic butyl methacrylate-based monolithic porous polymer was copolymerized with an ionizable monomer, [2-(methacryloyloxy)ethyl] trimethylammonium chloride to form a polymer monolith for solid phase extraction that also sustains anodal electroosmotic flow. Exposure of the PolyE-323 coating to the monolith forming mixture affected the performance of the chip by a minor amount; electrokinetic migration times increased by ∼5%, and plate numbers were reduced by an average of 5% for proteins and peptides. 1-mm long on-chip monolithic solid phase extraction columns showed reproducible, linear calibration curves (R2=0.9978) between 0.1 and 5nM BODIPY at fixed preconcentration times, with a capacity of 2.4pmol or 0.92mmol/L of monolithic column for cytochrome c. Solution phase on-bed trypsin digestion was conducted by capturing model protein samples onto the monolithic polymer bed. Complete digestion of the proteins was recorded for a 30min stop flow digestion, with high sequence coverage (88% for cytochrome c and 56% for BSA) and minimal trypsin autodigestion product. The polycationic coating and the polymer monolith materials proved to be compatible with each other, providing a high quality solid phase extraction bed and a robust coating to reduce protein adsorption and generate anodal flow, which is advantageous for electrospray. © 2011 Elsevier B.V.
Publication date
In
  • Journal of Chromatography A 1218, no. 26 (2011): 4039–4044.
LanguageEnglish
Peer reviewedYes
NPARC number21271526
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