高宗晔;史策;姚善泾;林东强

• 1:浙江大学生物质化工教育部重点实验室，浙江大学 化学工程与生物工程学院，浙江 杭州 310027

摘要(Abstract)：

针对单抗生产下游过程的瓶颈-蛋白A亲和分离，以提高产率和降低成本为目标，探讨了双柱连续流层析的新方法。从过程设计的关键因素出发，比较和分析了保留时间和蛋白浓度的影响，优化了分离条件，并应用于细胞培养液中单抗的分离。以两种蛋白A亲和介质MabSelect SuRe和UniMab为典型介质，考察了不同保留时间和IgG浓度下的穿透曲线，比较了保留时间和蛋白浓度对产率、介质利用率和缓冲液消耗的影响。结果表明，与常规的批次层析相比，双柱连续流层析拥有更优越的分离性能，低保留时间条件下的产率和介质利用率显著提高，缓冲液消耗减少，特别适合于高流速操作。对于从细胞培养液中分离单抗，双柱连续流层析的分离性能与常规批次层析相当，单抗纯度在97%以上、收率87%以上、聚集体含量低于2%、宿主细胞蛋白含量低于300 ppm，优势是产率提高，介质利用率从20%³0%提高到70%30%提高到70%⁸0%，，缓冲液消耗下降为常规批次层析过程的1/580%，，缓冲液消耗下降为常规批次层析过程的1/5¹/4，显示出良好的应用前景。

关键词(KeyWords)： 连续流层析;抗体;分离;产率;介质利用率

Abstract:

Keywords:

基金项目(Foundation): 国际科技合作专项项目（2015DFG42070）；国家自然科学基金（21776243，21476198）。

作者(Authors): 高宗晔;史策;姚善泾;林东强

DOI: 110.3969/j.issn.1003-9015.2018.00.019

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