[Analytical Chemistry] Thiol-ene monolithic pepsin microreactor with a 3D-printed interface for effi
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 enzym 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.
Alexander Jönsson, Rasmus R. Svejdal, Nanna B. Bøgelund, Tam T. T. N. Nguyen, Henrik Flindt, Jörg P. Kutter, Kasper D. Rand, and Josiane Penelope Lafleur Anal. Chem., Just Accepted Manuscript DOI: 10.1021/acs.analchem.6b05103 Publication Date (Web): March 21, 2017 Copyright © 2017 American Chemical Society