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Poly(ethylene glycol)‐based monolithic capillary columns for hydrophobic interaction chromatography of immunoglobulin G subclasses and variants
Author(s) -
Desire Christopher T.,
Arrua R. Dario,
Talebi Mohammad,
Lacher Nathan A.,
Hilder Emily F.
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300558
Subject(s) - monolith , chromatography , chemistry , ethylene glycol , polymerization , monolithic hplc column , monomer , resolution (logic) , hydrophilic interaction chromatography , peg ratio , polymer , high performance liquid chromatography , organic chemistry , catalysis , finance , artificial intelligence , computer science , economics
Polymer monoliths were prepared in 150 μm id capillaries by thermally initiated polymerization of PEG diacrylate for rapid hydrophobic interaction chromatography of immunoglobulin G ( I g G ) 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 I g G subclasses and variants, including mixtures of the κ variants of I g G 1, I g G 2, and I g G 3 as well as the κ and λ variants associated with I g G 1 and I g G 2. 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 κ variants of I g G 1, I g G 2, I g G 3, and I g G 4 but also for the κ and λ variants of I g G 1 and I g G 2. 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.