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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。
Explore the use of Schiff’s base method in the coupling of protein A ligands on the surface of super-macroporous polymer microspheres Preparation of affinity chromatography medium to hydrophilized polyacrylate macroporous microspheres as matrix, investigated the oxidant (H5IO6) on the amount of ligand conjugate. The surface morphology of the microspheres was characterized by scanning electron microscopy. The results showed that the microspheres could still retain its macropore structure after coupling Protein A. The dynamic loading of this kind of affinity media at different operating flow rates (361 ~ 3 600 cm / h) was investigated. Under the operating flow rate of 3 600 cm / h, the human antibody load decreased only by 10 %, Indicating that this kind of medium is suitable for rapid mass transfer of antibody macromolecules, which can meet the demand of rapid, efficient and high-throughput separation and purification.