基于超大孔聚合物微球的混合色谱模式层析介质的制备

Preparation of Mix-mode Chromatographic Supports Based on Gigaporous Polymer Microspheres

以甲基丙烯酸缩水甘油酯与乙二醇二甲基丙烯酸酯共聚的超大孔聚合物微球为基质,采用聚乙烯亚胺和丁基缩水甘油醚先后衍生微球的表面,制备成兼具阴离子交换与疏水相互作用的混合色谱模式层析介质。考察离子交换基团、疏水配基密度对蛋白载量、回收率的影响,结果表明,在离子交换容量0.2~0.5 mol/mL范围内,随着介质离子交换容量的增大,蛋白载量及回收率均呈增加趋势,蛋白载量最高值达40 mg/mL,回收率大于90%。当疏水配基的密度大于0.03 mol/mL时,介质开始表现疏水相互作用。此超大孔混合模式色谱介质在大于2000 cm/h的流速下依然能保持低于2 MPa的柱背压,同时在高流速下（2880 cm/h）纯化人血清中的抗体应用中表现良好的分辨率。此介质在高通量分离纯化应用方面具有巨大潜力。

The mix-mode chromatography supports were prepared, including anion exchange and hydrophobic interaction chromatography. This matrix was based on the gigaporous polymer microspheres from copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate. The microspheres were modified by poly （ ethylene imine） and butyl glycidyl ether for mix-mode chromatography. The effects of ion capacity and hydrophobic ligand density on protein capacity and recovery were evaluated. The result indicated that the protein capacity and recovery increased with ion capacity of the supports in the range of 0. 2-0. 5 mmol/mL. The max capacity was 40 mg/mL and the protein recovery could reach more than 90%. The hydrophobic interaction chromatography occurred when hydrophobic ligand density was more than 0. 03 mmol/mL. The back pressure on these supports was less than 2 MPa at 2000 cm/h. In the same time, the high resolution could be retained even at high flow rate of 2880 cm/h for purification of IgG from human serum. Therefore, this support showed a large potential in high through-put separation.