Immobilization of trypsin onto porous methacrylate-based monolith for flow-through protein digestion and its potential application to chiral separation using liquid chromatography

Monolithic columns for analytical applications have attracted the researcher's attention. In this work, laboratory-made organic-polymer monolithic column is modified with trypsin and further applied as a nanobiocatalyst microreactor stationary phase separating chiral compounds by liquid chromatography. The monolith was synthesized in-situ copolymerization of glycidyl methacrylate (GMA) ethylene glycol dimethacrylate (EDMA) or trimethylolpropane trimethacrylate (TRIM) crosslinking agent, porogen 1,4-butanediol/propanol/water (4:7:1 v/v) AIBN radical polymerization initiator inside PEEK silicosteel tubings (1.0 mm i.d × 100 mm) at 60 °C 12 h. A total monomer ratio (%T) agent (%C) 40:25 28:12 were to prepare poly-(GMA-co-EDMA) poly-(GMA-co-TRIM), respectively. produced monoliths introducing (10 mg/L) through ring-opening reaction epoxide group existing in column. trypsin-immobilized online/flow-through rapid digestion ?-casein sample into its peptide fragments. poly-(GMA-co-TRIM) has potential application be used HPLC separation R/S-citronellal enantiomers.