Preparation of polymeric monoliths by copolymerization of acrylate monomers with amine functionalities for anion-exchange capillary liquid chromatography of proteins
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Abstract
Two novel polymeric monoliths for anion-exchange capillary liquid chromatography of
proteins were prepared in a single step by a simple photoinitiated copolymerization
of 2-(diethylamino)ethyl methacrylate and polyethylene glycol diacrylate (PEGDA),
or copolymerization of 2-(acryloyloxy)ethyl trimethylammonium chloride and PEGDA,
in the presence of selected porogens. The resulting monoliths contained functionalities
of diethylaminoethyl (DEAE) as a weak anion-exchanger and quaternary amine as a strong
anion-exchanger, respectively. An alternative weak anion-exchange monolith with DEAE
functionalities was also synthesized by chemical modification after photoinitiated
copolymerization of glycidyl methacrylate (GMA) and PEGDA. Important physical and
chromatographic properties of the synthesized monoliths were characterized. The dynamic
binding capacities of the three monoliths (24 mg/mL, 56 mg/mL and 32 mg/mL of column
volume, respectively) were comparable or superior to values that have been reported
for various other monoliths. Chromatographic performance was also similar to that
provided by a modified poly(GMA-ethylene glycol dimethacrylate) monolith. Separation
of standard proteins was achieved under gradient elution conditions using these monolithic
columns. Peak capacities of 34, 58 and 36 proteins were obtained with analysis times
of 20-30 min. This work represents a successful attempt to prepare functionalized
monoliths via direct copolymerization of monomers with desired functionalities. Compared
to earlier publications, additional surface modifications were avoided and the PEGDA
crosslinker helped to improve the biocompatibility of the monolithic backbone.