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High temperature liquid chromatography of intact proteins using organic polymer monoliths and alternative solvent systems

journal contribution
posted on 2023-05-17, 02:30 authored by Causon, TJ, Nordborg, AMM, Robert ShellieRobert Shellie, Emily HilderEmily Hilder
Alternative approaches to conventional acetonitrile gradient methods for reversed-phase liquid chromatographic analysis of intact proteins have been investigated using commercial poly(styrene-co-divinylbenzene) monolithic columns (Dionex ProSwift™ RP-2H and RP-4H). Alternative solvents to acetonitrile (2-propanol and methanol) coupled with elevated temperatures demonstrated complementary approaches to adjusting separation selectivity and reducing organic solvent consumption. Measurements of peak area at increasing isothermal temperature intervals indicated that only minor (<5%) decreases in detectable protein recovery occurred between 40 and 100 °C on the timescale of separation (2–5 min). The reduced viscosity of a 2-propanol/water eluent at elevated temperatures permitted coupling of three columns to increase peak production (peaks/min) by 16.5%. Finally, narrow-bore (1 mm i.d.) columns were found to provide a more suitable avenue to fast, high temperature (up to 140 °C) separations.

Funding

Australian Research Council

History

Publication title

The Journal of Chromatography A

Volume

1217

Issue

21

Pagination

3519-3524

ISSN

0021-9673

Department/School

School of Natural Sciences

Publisher

Elsevier Science Bv

Place of publication

Po Box 211, Amsterdam, Netherlands, 1000 Ae

Rights statement

The definitive version is available at http://www.sciencedirect.com

Repository Status

  • Restricted

Socio-economic Objectives

Expanding knowledge in the chemical sciences

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