Development and evaluation of silica-based lectin microcolumns for glycoform analysis of alpha₁-acid glycoprotein.

ABSTRACT: Silica-based lectin microcolumns were developed and optimized for the separation and analysis of glycoform fractions in alpha₁-acid glycoprotein (AGP) based on both the degree of branching and level of fucosylation. Concanavalin A (Con A) and Aleuria Aurantia lectin (AAL) were immobilized onto HPLC-grade silica by reductive amination and packed into 2.1 mm i.d. × 5.0 cm microcolumns. Factors examined for these microcolumns include their protein content, binding capacity, binding strength and band-broadening under isocratic conditions (Con A) or step elution conditions (AAL) and in the presence of various flow rates or temperatures. These factors were examined by using experiments based on frontal analysis, zonal elution, peak profiling and peak decay analysis. Up to 200 μg AGP c

SUBMITTER: Zhang C 

PROVIDER: S-EPMC6668930 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature