应用于抗体分离纯化的疏水电荷诱导色谱配基和性能评价研究.pdfVIP

应用于抗体分离纯化的疏水电荷诱导色谱配基和性能评价 史清洪，程征，孙舒，孙彦 天津大学化工学院生物化工系，天津，300072 摘要：获取高品质（高纯度、低杂质）的抗体产品已经成为抗体制备中的一个瓶颈问题。本文对新型疏水电荷 诱导色谱（HCIC ）配基 4 （咪唑-1-基）苯胺在抗体吸附和分离中的性能进行了研究。抗体和血清白蛋白（BSA, 抗体生产中的重要杂蛋白）在烯丙基活化法制备得到的 HCIC 色谱吸附剂 AN-Sepharose 上的吸附平衡结果显 示，在 20 mmol/L 磷酸缓冲液（pH 8.0 ）中，抗体在AN-Sepharose 上的吸附展现出一定的耐盐特性，同时这种 吸附受到乙二醇的抑制；而在氯化钠存在条件下 BSA 的吸附则显著不同于抗体。随着缓冲液pH 值降低，抗体 的吸附容量迅速减小，意味着大部分抗体可以通过调节流动相的pH 值加以洗脱。从蛋白质混合物中纯化抗体的 结果显示，抗体的纯度超过90% 。由此，该色谱介质显示了其在抗体分离中的巨大应用潜力。 关键词：抗体；疏水电荷诱导色谱；蛋白吸附；色谱配基；血清白蛋白 中图分类号： 文献标识码：A 文章编号： Performance of a novel hydrophobic charge induction chromatographic bead in antibody purification SHI Qinghong, CHEN Zheng, SUN Shu, SUN Yan (Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China) Abstract: In the last two decades, the issues in the antibody purification have become worse due to the titer increase in cell culture. Now the industry is beginning to look for higher affinity columns and faster beads to increase throughput. By virtual screening technique, 4-(1H-imidazol-1-yl) aniline was obtained from commercial chemical database and was coupled onto commercial agarose gel, Sepharose CL-6B, through allyl chemistry to fabricate chromatographic bead, AN-Sepharose, for hydrophobic charge induction chromatography (HCIC). The adsorption of antibody and bovine serum albumin (BSA) on AN-Sepharose was investigated in batch experiment and chromatographic column respectively. The results indicated that adsorption of antibody on AN-Sepharose had a typical salt resistance and partially inhibited by ethyl glycol whereas BSA adsorption on AN-Sepharose had a complete difference