Synthesis of Boronic Acid-functionalized Soluble Dendrimers and its Application in Detection of Human Liver Microsomal Glycoprotein Based on Enzyme-linked Immunosorbent Assay
Abstract It is difficult for analysis of glycoproteins in the biological sample due to the low abundance, susceptiblility to be inhibited and interfered by other non-glycoproteins. An enrichment step is usually required before glycoprotein analysis, but the operation steps of conventional solid-phase based glycoprotein enrichment methods are difficult to be compatible with the most classical enzyme-linked immuno sorbent assay (ELISA) technique. In this study, a novel water-soluble dendrimer based boronic acid capture (DBC) method was developed using PAMAM 4.0 as carrier and boronic acid as affinity group. The method was applied to detection of glycoproteins in human liver microsomes using ELISA. The DBC enrichment conditions were optimized by model glycoprotein, and then its sensitivity and anti-interference ability were investigated. The optimized method was applied to enrich glycoproteins in human liver microsomal. The results showed that the enrichment selectivity of DBC for glycoprotein could be up to 100,000 folds, and the signal of glycoprotein could be improved by 100 times. In conclusion, using DBC as a novel enrichment material for glycoprotein, combined with ELISA analysis, high sensitivity and selectivity detection could be achieved in biological samples with only one simple incubation step, which makes it a useful tool for glycoprotein research.