Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses

Alexander Jönsson; Rasmus R Svejdal; Nanna Bøgelund; Tam T T N Nguyen; Henrik Flindt; Jörg P Kutter; Kasper D Rand; Josiane P Lafleur

doi:10.1021/acs.analchem.6b05103

Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses

Alexander Jönsson, Rasmus R Svejdal, Nanna Bøgelund, Tam T T N Nguyen, Henrik Flindt, Jörg P Kutter, Kasper D Rand, Josiane P Lafleur

30 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Analytical Chemistry
Vol/bind	89
Udgave nummer	8
Sider (fra-til)	4573-4580
Antal sider	8
ISSN	0003-2700
DOI	https://doi.org/10.1021/acs.analchem.6b05103
Status	Udgivet - 18 apr. 2017

Adgang til dokumentet

10.1021/acs.analchem.6b05103Licens: CC BY-ND

acs.analchemForlagets udgivne version, 2,35 MB

Citationsformater

@article{f9972df2c07544109d6e0723b28610d2,

title = "Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses",

abstract = "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.",

keywords = "Journal Article",

author = "Alexander J{\"o}nsson and Svejdal, {Rasmus R} and Nanna B{\o}gelund and Nguyen, {Tam T T N} and Henrik Flindt and Kutter, {J{\"o}rg P} and Rand, {Kasper D} and Lafleur, {Josiane P}",

year = "2017",

month = apr,

day = "18",

doi = "10.1021/acs.analchem.6b05103",

language = "English",

volume = "89",

pages = "4573--4580",

journal = "Analytical Chemistry",

issn = "0003-2700",

publisher = "American Chemical Society",

number = "8",

}

TY - JOUR

T1 - Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses

AU - Jönsson, Alexander

AU - Svejdal, Rasmus R

AU - Bøgelund, Nanna

AU - Nguyen, Tam T T N

AU - Flindt, Henrik

AU - Kutter, Jörg P

AU - Rand, Kasper D

AU - Lafleur, Josiane P

PY - 2017/4/18

Y1 - 2017/4/18

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1021/acs.analchem.6b05103

DO - 10.1021/acs.analchem.6b05103

M3 - Journal article

C2 - 28322047

SN - 0003-2700

VL - 89

SP - 4573

EP - 4580

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 8

ER -

Thiol-ene Monolithic Pepsin Microreactor with a 3D-Printed Interface for Efficient UPLC-MS Peptide Mapping Analyses

Abstract

Adgang til dokumentet

Fingeraftryk

Citationsformater