超大孔聚合物亲和色谱层析介质的制备及评价

发布时间：2018-01-12 23:26

本文关键词：超大孔聚合物亲和色谱层析介质的制备及评价　出处：《分析测试学报》2016年12期 　论文类型：期刊论文

【摘要】：探索了用席夫碱法在超大孔聚合物微球表面偶联Protein A配基制备亲和层析介质的方法,以亲水化后的聚丙烯酸酯类超大孔微球为基质,考察了氧化剂浓度(H5IO6)对配基偶联量的影响,以扫描电子显微镜表征微球表面形貌,结果发现其偶联Protein A后微球仍能够保持其大孔结构。考察了该类亲和介质在不同操作流速(361~3 600 cm/h)下的动态载量,在3 600 cm/h操作流速下,人抗体载量与361 cm/h相比仅下降10%左右,表明该类介质适合抗体大分子的快速传质,能够满足快速、高效、高通量分离纯化的需求。
[Abstract]:An affinity chromatography medium was prepared by coupling Protein A ligand on the surface of macroporous polymer microspheres by Schiff base method. The hydrophilic macroporous polyacrylate microspheres were used as the matrix. The effect of oxidant concentration (H 5IO 6) on ligand coupling amount was investigated. The surface morphology of microspheres was characterized by scanning electron microscope (SEM). The results showed that the microspheres could maintain their macroporous structure after coupling with Protein A. the dynamic loading capacity of this kind of affinity medium was investigated at different operating velocities of 361n 3600 cm / h. At the operating flow rate of 3 600 cm/h, the human antibody load decreased by only about 10% compared with 361 cm/h, which indicated that the medium was suitable for rapid mass transfer of antibody macromolecules and could satisfy the requirement of rapid mass transfer. High efficiency, high throughput separation and purification requirements.
【作者单位】：北京石油化工学院化学工程学院;中国科学院过程工程研究所生化工程国家重点实验室;
【基金】：北京市自然科学基金(2162013) 国家自然科学基金(51103158) 北京石油化工学院优秀人才培育计划项目(080318620081/037) 北京高等学校高水平人才交叉培养“实培计划”项目(201507,201506) 2016大学生URT项目(16032082001/009)
【分类号】：O658.1
【正文快照】： 生化工程国家重点实验室,北京100190)亲和层析是利用生物活性物质之间的特异亲和力,使目标产物得以分离纯化的液相层析方法,可应用于任何两种有特异性相互作用的生物大分子[1]。由于抗体与抗原作用具有高度的专一性,且该亲和力极强,亲和层析技术兼具高收率、高纯度、能保持生

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