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

Citation

Causon, TJ and Nordborg, AMM and Shellie, RA and Hilder, EF, High temperature liquid chromatography of intact proteins using organic polymer monoliths and alternative solvent systems, The Journal of Chromatography A, 1217, (21) pp. 3519-3524. ISSN 0021-9673 (2010) [Refereed Article]


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DOI: doi:10.1016/j.chroma.2010.03.033

Abstract

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.

Item Details

Item Type:Refereed Article
Keywords:High temperature liquid chromatography; Polymer monoliths; Proteins
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Separation Science
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Chemical Sciences
Author:Causon, TJ (Mr Timothy Causon)
Author:Nordborg, AMM (Dr Anna Nordborg)
Author:Shellie, RA (Associate Professor Robert Shellie)
Author:Hilder, EF (Professor Emily Hilder)
ID Code:63381
Year Published:2010
Funding Support:Australian Research Council (DP0987318)
Web of Science® Times Cited:15
Deposited By:Austn Centre for Research in Separation Science
Deposited On:2010-05-03
Last Modified:2015-01-21
Downloads:0

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