Polyethylene glycol diacrylate-based supermacroporous monolithic cryogel as high-performance liquid chromatography stationary phase for protein and polymeric nanoparticle separation

A supermacroporous monolithic cryogel was directly prepared by in situ cryo-copolymerization in a stainless steel cartridge (70 mm × 5.0 mm I.D.) using methacrylic acid (MAA) as functional monomer and polyethylene glycol diacrylate (PEGDA) as crosslinker. The highly crosslinked (90%, molar ratio) poly(MAA–PEGDA) cryogel had more uniform supermacropores with a mean diameter of 25 μm compared to the poly(acrylamide)-based cryogels. The viability of poly(MAA–PEGDA) cryogel as a medium was demonstrated for separations of lysozyme from chicken egg white (CEW) and water-soluble poly(N-isopropylacrylamide-co-3-(dimethylamino) propyl methacrylamide) (NIPAM-DMAPMA) nanoparticles from its crude reaction solution. The dynamic binding capacities of lysozyme and the polymeric nanoparticles were 4.51 × 10−3 μmol/ml and 33.4 μg/ml, respectively. The lysozyme recovered from the above separations had a purity of more than 85%, and retained 90% of its enzymatic activity.