Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses.
Alexander JönssonRasmus R SvejdalNanna BøgelundTam T T N NguyenHenrik FlindtJörg P KutterKasper Dyrberg RandJosiane P LafleurPublished in: Analytical chemistry (2017)
To improve the sample handling, and reduce cost and preparation time, of peptide mapping LC-MS workflows in protein analytical research, we here investigate the possibility of replacing conventional enzymatic digestion methods with a polymer microfluidic chip based enzyme reactor. Off-stoichiometric thiol-ene is utilized as both bulk material and as a monolithic stationary phase for immobilization of the proteolytic enzyme pepsin. The digestion efficiency of the, thiol-ene based, immobilized enzyme reactor (IMER) is compared to that of a conventional, agarose packed bed, pepsin IMER column commonly used in LC-MS based protein analyses. The chip IMER is found to rival the conventional column in terms of digestion efficiency at comparable residence time and, using a 3D-printed interface, be directly interfaceable with LC-MS.
Keyphrases
- liquid chromatography
- mass spectrometry
- anaerobic digestion
- high throughput
- simultaneous determination
- high resolution
- solid phase extraction
- capillary electrophoresis
- circulating tumor cells
- tandem mass spectrometry
- molecularly imprinted
- wastewater treatment
- ionic liquid
- protein protein
- high density
- hydrogen peroxide
- binding protein
- magnetic nanoparticles