人源化抗人TNFα单克隆抗体纯化工艺的优化

发布时间：2018-01-16 16:21

本文关键词：人源化抗人TNFα单克隆抗体纯化工艺的优化　出处：《沈阳药科大学学报》2017年01期 　论文类型：期刊论文

【摘要】：目的考察中国仓鼠卵巢细胞(Chinese hamster ovary,CHO细胞)表达的人源化抗人TNFα单克隆抗体的亲和色谱过程中的杂质洗脱工艺,优化目的蛋白洗脱工艺条件。方法以目的蛋白回收率和宿主细胞蛋白残留量为评价指标,筛选最佳的杂质洗脱缓冲液;利用实验设计方法,以纯度、洗脱峰pH值、宿主细胞蛋白残留量为评价指标,考察目的蛋白洗脱缓冲液的pH值和浓度对工艺的影响,优化目的蛋白洗脱工艺条件。结果优化后的亲和色谱工艺,在保持目的蛋白高回收率(优化前为86.5%,优化后为86.7%)的同时,可显著降低目的蛋白洗脱峰中宿主细胞蛋白残留量(由1 218×10-6降低至78×10-6),提高目的蛋白纯度(优化前为95.5%,优化后为96.6%),减轻后续纯化压力;提高了目的蛋白洗脱峰的pH值(由3.59提高到4.25),有利于目的蛋白的稳定。结论优化了人源化抗人TNFα单克隆抗体的亲和色谱工艺,本工艺有效、可行。
[Abstract]:Objective to investigate the China hamster ovary cells (Chinese hamster ovary, CHO cells) expression of humanized alpha impurities of anti human TNF monoclonal antibody affinity chromatography in the elution process, optimize elution conditions. Methods to recovery of protein and host cell protein residues as the evaluation index, select the best impurity elution buffer the use of liquid; experimental design method, the purity of elution peak value of pH, host cell protein residues as the evaluation index, the effects of elution buffer pH and concentration of technology, optimization objective protein elution conditions. Affinity chromatography process optimized results, while maintaining the target protein with high recovery rate (optimization of 86.5% before and after optimization is 86.7%) at the same time, can significantly reduce the elution of residual host cell protein peaks (from 1218 down to 78 * 10-6 * 10-6), to improve the purity of target protein (before optimization is 95.5%, after optimization is 96.6%), reduce the pressure to improve subsequent purification; protein elution peak value of pH (from 3.59 to 4.25), is favorable to protein stability. Conclusion the affinity chromatography process optimization of humanized monoclonal antibody against human TNF, this technology is effective and feasible.

【作者单位】：沈阳药科大学生命科学与生物制药学院;沈阳三生制药有限责任公司;
【分类号】：R392.11
【正文快照】： 肿瘤坏死因子α(tumor necrosis factor-alpha,TNFα)是与类风湿性关节炎、克罗恩病、银屑病及强直性脊髓炎等多种自身免疫病密切相关的重要炎症因子[1-2],人肿瘤坏死因子α(h TNFα)中和抗体被认为是目前治疗这些疾病最有效的药物之一[3]。典型的单克隆抗体纯化方法一般为三

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