Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses.
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.
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Alexander
A
Jönsson
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Rasmus R
RR
Svejdal
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Nanna
N
Bøgelund
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Tam T T N
TTTN
Nguyen
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Henrik
H
Flindt
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Jörg P
JP
Kutter
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Kasper D
0000-0002-6337-5489
KD
Rand
Department of Pharmacy, Copenhagen University , Universitetsparken 2, Copenhagen E DK-2100, Denmark.
Josiane P
0000-0003-4459-8801
JP
Lafleur
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004800:Enzymes, Immobilized; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D056656:Lab-On-A-Chip Devices; D013058:Mass Spectrometry; D010434:Pepsin A / Q000737:chemistry / Q000378:metabolism*; D010449:Peptide Mapping / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000032:analysis* / Q000378:metabolism; D011108:Polymers / Q000737:chemistry; D066330:Printing, Three-Dimensional; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D004800:Enzymes, Immobilized; D006454:Hemoglobins; D010455:Peptides; D011108:Polymers; D013438:Sulfhydryl Compounds; D010434:Pepsin A