Synthesis and application of a macroporous boronate affinity monolithic column using a metal-organic gel as a porogenic template for the specific capture of glycoproteins

• Published in: Journal of Chromatography A Vol. 1218; no. 51; pp. 9194 - 9201
• Main Authors: Yang, Fan, Lin, Zian, He, Xiwen, Chen, Langxing, Zhang, Yukui
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 23.12.2011; Elsevier
• Subjects: Acrylic Resins - chemistry; Affinity; Analytical chemistry; Analytical, structural and metabolic biochemistry; Animals; Binding; Biological and medical sciences; Boronate affinity chromatography; Boronic Acids - chemistry; Catechols - analysis; Cattle; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography; Chromatography, Affinity - instrumentation; Chromatography, Affinity - methods; Chromatography, High Pressure Liquid; Dynamic tests; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Glycoprotein; Glycoproteins; Glycoproteins - isolation & purification; Horseradish Peroxidase - isolation & purification; Hydroquinones - analysis; Intrusion; Macroporous monolithic column; Metal-organic gels; Microscopy, Electron, Scanning; Monomers; Other chromatographic methods; Porosity; Pressure; Proteins; Scanning electron microscopy; Temperature; Transferrin - isolation & purification; Glycoproteins; Macroporous monolithic column; Boronate affinity chromatography; Metal-organic gels; Transferrin; In situ; Acrylamide derivative copolymer; Porous material; Blood; Surface structure; Affinity adsorbent; Characterization; Crosslinked copolymer; Separation method; Binding capacity; Affinity chromatography; Ungulata; Ferroprotein; Scanning electron microscopy; Iron III Nitrates Hydrates; Bovine; Monolithic column; Glycoprotein; Protein; Boron copolymer; Mercury porosimetry; Vertebrata; Mammalia; Animal; Morphology; Preparation; Artiodactyla; Methacrylate copolymer; Radical copolymerization; Blood serum; Physicochemical properties; Macroporosity
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2011.10.049

► An alternative method was developed for the synthesis of a macroporous boronate affinity monolithic column in a convenient way using MOG as a porogenic template. ► The monolithic column had the advantage of good macropore distribution and permeability for further high-throughput and efficient separations of glycoproteins. ► A one-step purification of transferrin from bovine serum sample was of success. A macroporous boronate affinity monolithic column was prepared and applied to specifically capture glycoproteins using metal-organic gels (MOGs) as a porogenic template. This newly explored application of MOGs has proven to be a more convenient method for the formation of macropores in contrast to traditional porogenic methods. The poly (3-acrylamidophenylboronic acid-co-ethylene dimethacrylate) monolithic columns were synthesized in stainless columns by in situ polymerization. To fabricate the macroporous formation with a uniformed open-channel network, the preparation conditions, such as reaction temperature, the concentration of the MOGs and the ratio of monomers were systematically investigated. The prepared macroporous monoliths were characterized by scanning electron microscope (SEM) and mercury intrusion porosimetry. Furthermore, horseradish peroxidase (HRP) and transferrin (TF) were chosen as test glycoproteins, and the chromatographic analysis demonstrated that the macroporous boronate affinity monoliths exhibited a higher selectivity and better dynamic binding capacity toward glycoproteins compared with non-glycoproteins. The resulted affinity monolithic column was successfully employed to specifically capture TF from a bovine serum sample.