Development of molecularly imprinted Acrylamide-Acrylamido phenylboronic acid copolymer microbeads for selective glycosaminoglycan separation in children urine

Background: in this study, we synthesized molecularly imprinted copolymers for liquid chromatography columns as a separator for glycosaminoglycan (dermatan sulfate; DS and chondroitin sulfate; CS) in urine. Materials and methods: Acrylamide and acrylamido phenylboronic acid were used as monomers, acrylamide was used for as base monomer to attract negatively charged groups and acrylamido phenylboronic acid (AAPBA) residues used to form diol bonds between sugar and boronic acid residues to strengthen the attraction. These monomers were synthesized by using precipitation polymerization to form uniform spheres, which are more durable for the pressurized chromatographic systems. Trimethylolpropane trimethacrylate and AIBN were used as crosslinker and starter, respectively. Results: These GAG selective polymers were filled by pressurized flow into the steel (4.6 mm x1.6 mm) columns, then imprinted GAGs were extracted and analyzed to calculate binding capacity of each milligram polymer. Calibration curves of the GAG selective columns were obtained 62.5 -1000 ng/mL less than 5% coefficient variation, and lower matrix effect. Conclusion: Our imprinted columns separated different GAGs from urine specifically and sensitively. Matrix effect was at an ignorable level thus the challenging use.