Preparation and evaluation of a phenylboronate affinity monolith for selective capture of glycoproteins by capillary liquid chromatography
Preparation and evaluation of a phenylboronate affinity monolith for selective capture of glycoproteins by capillary liquid chromatography
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用于毛细管液相色谱选择性捕获糖蛋白的苯硼酸亲和整体料的制备和评价
DOI：
10.1039/c1an15180k
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发表时间：
2011-01-01
期刊：
影响因子：
4.2
通讯作者：
Chen, Guo Nan
中科院分区：
文献类型：
--
作者：
Lin, Zi An;Pang, Ji Lei;Chen, Guo Nan
制备了一种苯硼酸盐亲和单体，并将其应用于从未分离的蛋白质混合物中选择性捕获糖蛋白。以4-乙烯基苯基硼酸（VPBA）为功能单体，二甲基丙烯酸乙酯（EDMA）为交联剂，二甘醇和乙二醇为二元致孔溶剂，偶氮二异丁腈（AIBN）为引发剂，在100 μ m的毛细管中通过原位聚合法制备了该单体。对制备的整体石进行了形貌、孔隙性质和识别性能的表征。以标准糖蛋白和非糖蛋白为模型蛋白，评价其选择性和动态结合能力。色谱结果表明，与非糖蛋白相比，苯硼酸盐亲和单体对糖蛋白具有更高的选择性和结合能力。将所得的苯硼酸亲和整体石作为固相微萃取（in-tube SPME）的吸附剂，并通过捕获蛋清样品中的卵清蛋白来评估整体石的萃取性能。
A phenylboronate affinity monolith was prepared and applied to the selective capture of glycoproteins from unfractionated protein mixtures. The monolith was synthesized in a 100 mu m i.d capillary by an in situ polymerization procedure using a pre-polymerization mixture consisting of 4-vinylphenylboronic acid (VPBA) as functional monomer, ethylene dimethacrylate (EDMA) as crosslinker, diethylene glycol and ethylene glycol as binary porogenic solvents, and azobisisobutyronitrile (AIBN) as initiator. The prepared monolith was characterized in terms of the morphology, pore property, and recognition property. The selectivity and dynamic binding capacity were evaluated by using standard glycoproteins and nonglycoproteins as model proteins. The chromatographic results demonstrated that the phenylboronate affinity monolith had higher selectivity and binding capacity for glycoprotein than nonglycoprotein. The resulting phenylboronate affinity monolith was used as the sorbent for in-tube solid phase microextraction (in-tube SPME), and the extraction performance of the monolith was assessed by capture of ovalbumin from egg white sample.