Polymermonoliths were prepared in 150 um id capillaries by thermally initiated polymerization of PEG diacrylate for rapid hydrophobic interaction chromatography of immunoglobulin G (IgG) subclasses and related variants. Using only one monomer in the polymerization mixture allowed ease of optimization and synthesis of the monolith. The performance of the monolith was demonstrated by baseline resolution of IgG subclasses and variants, including mixtures of the k variants of IgG1, IgG2, and IgG3 as well as the k and ? variants associated with IgG1 and IgG2. The effect of eluent concentration and pH on the separation efficiency of studied proteins was also explored, allowing almost baseline resolution to be achieved for mixtures of the k variants of IgG1, IgG2, IgG3, and IgG4 but also for the k and ? variants of IgG1 and IgG2. The results showed significant improvement in the separations in terms of the tradeoff between analysis time and resolution, while maintaining a simple methodology, in comparison to previous reports. The synthesized monolith was also used for the separation of isoforms of a therapeutic monoclonal antibody.

Item Type:	Refereed Article
Keywords:	hydrophobic interaction chromatography; immunoglobulin G; proteins; polymer monoliths
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
UTAS Author:	Desire, CT (Mr Christopher Desire)
UTAS Author:	Arrua, RD (Dr Dario Arrua)
UTAS Author:	Talebi, M (Dr Mohammad Talebi)
UTAS Author:	Hilder, EF (Professor Emily Hilder)
ID Code:	86948
Year Published:	2013
Web of Science® Times Cited:	19
Deposited By:	Austn Centre for Research in Separation Science
Deposited On:	2013-11-04
Last Modified:	2022-08-30
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