A cross-linked polymer possessing a high density of hydrazide groups: high-throughput glycan purification and labeling for high-performance liquid chromatography analysis

Previously, we communicated that rapid and efficient purification of carbohydrates can be achieved by employing sugar-specific chemical ligation with hydrazide-functionalized polymer beads; we termed this process ‘glycoblotting’. The polymer beads are designed to recover carbohydrates by an imine exchange reaction in which hydrazone bonds between the beads and carbohydrates are transferred to oxime bonds between the aminooxy probe and the carbohydrates. To apply our concept to reductive amination with common fluorescent dyes, such as 2-aminobenzamide, the method for releasing the carbohydrates from the beads was examined, and we found that heating the beads with several percentages of acetic acid was efficient. Additionally, we obtained fundamental data on our novel method, such as the recovery ratio, the quantitative capability and the reproducibility. From the results, we concluded that rapid and accurate glycan analysis can be achieved with this novel method. Overall, these novel technologies represent a significant advance toward more efficient glycan analyses, especially by using high-performance liquid chromatography.