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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 HilderAlternative 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 AVolume
1217Issue
21Pagination
3519-3524ISSN
0021-9673Department/School
School of Natural SciencesPublisher
Elsevier Science BvPlace of publication
Po Box 211, Amsterdam, Netherlands, 1000 AeRights statement
The definitive version is available at http://www.sciencedirect.comRepository Status
- Restricted