Boronic acid functionalized Fe3O4 magnetic microspheres for the specific enrichment of glycoproteins

Glycoproteins are useful biomarkers and therapeutic targets for a number of diseases, including infections and cancer. However, identification and isolation of low-abundant glycoproteins remains a significant challenge that limits their application. Thus, methods of specific and selective glycoprotein enrichment are required. In this study, novel phenylboronic acid functionalized magnetic microspheres were successfully synthesized. Fe3O4 microspheres were synthesized by using a hydrothermal method and were coated with tetraethyl orthosilicate using an ultrasonic method to form a core-shell structure. Compared to the conventional mechanical stirring for 12 h, the ultrasonic method saved about 7 h in processing time, and the home-made magnetic microspheres had better dispersibility and homogeneity. Subsequently, the magnetic microspheres were modified by addition of an amino group and a carboxyl group, in sequence. Finally, 3-aminophenylboronic acid, as the functional monomer, was linked to the magnetic microspheres for capturing glycoprotein/glycopeptides. The results of this study indicate that phenylboronic acid functionalized magnetic microspheres show excellent adsorption performance toward glycoprotein/glycopeptides. The maximum absorbing capacity of the microspheres for fetuin was 108 mg/g, and the enrichment efficiency reached 89.7%, indicating their potential to separate and enrich glycoproteins from the complex biological samples.