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FlashPack: Fast and simple preparation of ultra-high performance capillary columns for LC-MS

Sergey Kovalchuk, Ole N. Jensen, Adelina Rogowska-Wrzesinska
doi: https://doi.org/10.1101/426676
Sergey Kovalchuk
1Department of Biochemistry & Molecular Biology and VILLUM Center for Bioanalytical Sciences, University of Southern Denmark, Campusvej 55, DK-5230, Odense M, Denmark
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Ole N. Jensen
1Department of Biochemistry & Molecular Biology and VILLUM Center for Bioanalytical Sciences, University of Southern Denmark, Campusvej 55, DK-5230, Odense M, Denmark
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Adelina Rogowska-Wrzesinska
1Department of Biochemistry & Molecular Biology and VILLUM Center for Bioanalytical Sciences, University of Southern Denmark, Campusvej 55, DK-5230, Odense M, Denmark
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  • For correspondence: adelinar@bmb.sdu.dk
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Abstract

Capillary ultra-high-pressure liquid chromatography (cUHPLC) is essential for in-depth characterization of complex biomolecule mixtures by LC-MS. We developed a simple and fast method called FlashPack for custom packing of capillary columns of 50-100 cm length with sub-2-μm sorbent particles. FlashPack uses high sorbent concentrations of 500-1000 mg/ml for packing at relatively low pressure of 100 bar. Column blocking by sorbent aggregation is avoided during the packing of sorbent particles by gentle mechanical tapping of the capillary proximal end by a slowly rotating magnet bar. Utilizing a standard 100 bar pressure bomb, Flashpack allows for production of 15-25 cm cUHPLC columns within a few minutes and of 50 cm cUHPLC columns in less than an hour. Columns exhibit excellent reproducibility of back-pressure, retention time and resolution (CV 8,7 %). FlashPack cUHPLC columns are inexpensive, robust and deliver performance comparable to commercially available cUHPLC columns. The FlashPack method is versatile and enables production of cUHPLC columns using a variety of sorbent materials.

Abbreviations
ACN
acetonitrile
BSA
bovine serum albumin
CAA
2-Chloroacetamide
cUHPLC
capillary ultra-high performance liquid chromatography
DB
database
EIC
extracted ion current chromatogram
ESI
electro-spray ionization
FA
formic acid
FDR
false discovery rate
FS
fused silica
FW
full width
FWHM
full width at half maximum
ID
internal diameter
LC
liquid chromatography
LC-MS
liquid chromatography-mass spectrometry
MQ
MaxQuant
OD
outer diameter
RT
retention time
SDC
Sodium deoxycholate
TCEP
Tris(2-carboxyethyl)phosphine
TEAB
triethylammonium bicarbonate
TFA
trifluoric acid
TIC
total ion current chromatogram
Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 27, 2018.
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FlashPack: Fast and simple preparation of ultra-high performance capillary columns for LC-MS
Sergey Kovalchuk, Ole N. Jensen, Adelina Rogowska-Wrzesinska
bioRxiv 426676; doi: https://doi.org/10.1101/426676
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FlashPack: Fast and simple preparation of ultra-high performance capillary columns for LC-MS
Sergey Kovalchuk, Ole N. Jensen, Adelina Rogowska-Wrzesinska
bioRxiv 426676; doi: https://doi.org/10.1101/426676

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