title: Preparation of boronate-functionalized molecularly imprinted monolithic column with polydopamine coating for glycoprotein recognition and enrichment.
abstract: A novel imprinting strategy using reversible covalent complexation of glycoprotein was described for creating glycoprotein-specific recognition cavities on boronate-functionalized monolithic column. Based on it, a molecularly imprinted monolithic column was prepared by self-polymerization of dopamine (DA) on the surface of 4-vinylphenylboronic acid (VPBA)-based polymeric skeletons after reversible immobilization of horseradish peroxidase (HRP). Due to the combination of boronate affinity and surface imprinting of DA, the stable and accessible recognition sites in the as-prepared imprinted monolith could be obtained after the removal of the template, which facilitated the rebinding of the template and provided good reproducibility and lifetime of use. The recognition behaviors of proteins on the bare VPBA-based, HRP-imprinted and nonimprinted monolithic columns were evaluated in detail and the results showed that the HRP-imprinted monolith exhibited higher recognition ability toward the template than another two monolithic columns. Not only nonglycoproteins but also glycoproteins can be well separated with the HRP-imprinted monolith. In addition, the feasibility of the HRP-imprinted monolith, adopted as an in-tube solid phase microextraction (in-tube SPME), was further assessed by selective extraction and enrichment of HRP from human serum. The good results demonstrated its potential in glycoproteome analysis.
authors.affiliation: Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian 350108, China. Electronic address: zianlin@fzu.edu.cn.
authors.forename: Zian
authors.initials: Z
authors.lastname: Lin
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Tan
authors.forename: Lixiang
authors.initials: L
authors.lastname: Sun
authors.forename: Ruifang
authors.initials: R
authors.lastname: Yu
authors.forename: Huanghao
authors.initials: H
authors.lastname: Yang
authors.forename: Guonan
authors.initials: G
authors.lastname: Chen
mesh_terms: D001897:Boronic Acids / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007211:Indoles / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D011108:Polymers / Q000737:chemistry*; D015203:Reproducibility of Results; D052617:Solid Phase Microextraction
chemical_list: D001897:Boronic Acids; D004800:Enzymes, Immobilized; D006023:Glycoproteins; D007211:Indoles; D011108:Polymers; C568283:polydopamine; D006735:Horseradish Peroxidase
keywords: Boronate affinity; Glycoprotein; Molecular imprinting; Monolithic column; Recognition