期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1319, 期 -, 页码 141-147出版社
ELSEVIER
DOI: 10.1016/j.chroma.2013.10.059
关键词
Molecular imprinting; Monolithic column; Recognition; Glycoprotein; Boronate affinity
资金
- National Natural Science Foundation of China [21005018, 21375018]
- National Basic Research Program of China [2010CB732403]
- National Natural Science Funds for Distinguished Young Scholar [21125524]
- Doctoral Fund of Ministry of Education [20103514120002]
- Program for Changjiang Scholars and Innovative Research Team in University [IRT1116]
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. (C) 2013 Elsevier B.V. All rights reserved.
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