Preparation of Novel magnetic boronic acid-affinity material and investigation of adsorption performance toward ovalbumin.

Glycoproteins play a crucial role in various biological processes, including protein folding, information transmission, nerve conduction, and molecular recognition. In this study, a novel magnetic phenylboronic acid-functionalized covalent organic framework core-shell material (Fe3O4@BBCOF-BA) was successfully synthesized for the highly selective enrichment of glycoproteins from complex biological samples. Systematic optimization of experimental conditions revealed that under weakly alkaline conditions at pH 8.5, the material specifically recognized glycoproteins through boronate ester bonds. The maximum adsorption capacity for ovalbumin (OVA) reached as high as 469.45 mg/g, representing a two-to threefold improvement over traditional magnetic boronate affinity materials. Adsorption kinetics indicated that equilibrium was reached within 120 min, and rapid separation was achieved within 30 s utilizing the superparamagnetic property. Mild elution was performed using an ethanol-formic acid (90:10, v/v) solution, achieving an OVA recovery rate exceeding 90 %. Selectivity experiments confirmed that the material exhibited significantly higher adsorption of glycoproteins (OVA, HRP) than non-glycoproteins (BSA, Lys). Practical application evaluation demonstrated efficient enrichment of trace glycoproteins from 500-fold diluted egg white and complex protein mixtures. SDS-PAGE analysis confirmed low non-specific adsorption, underscoring its strong potential for application in the pretreatment of complex biological samples.