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    • 7. 发明公开
    • ANTIBODY PURIFICATION AND PURITY MONITORING
    • 抗体纯化和纯度监测
    • EP2970865A1
    • 2016-01-20
    • EP14763076.8
    • 2014-03-17
    • Alder Biopharmaceuticals, Inc.
    • LATHAM, John, A.GARCIA-MARTINEZ, Leon, F.ALLISON, Daniel, S.DAVIN, Steven, D.DO, Hoa, BinhLEE, Geoffrey, F.OJALA, Ethan, W.YOUNG, Mark
    • C12N1/19
    • Processes for producing and purifying recombinant proteins are disclosed. In particular, the present disclosure provides processes of producing and purifying multi-subunit proteins expressed in yeast or filamentous fungal cells. The production and/or purification of such proteins are monitored for impurities, preferably using lectin binding assays, such that one or more process parameters may be adjusted to maximize the amount of desired recombinant protein and minimize the amount of glycosylated impurities. The processes can also be monitored for other undesired product-associated impurities, such as aggregates and nucleic acids. In exemplary embodiments, the recombinant proteins are multi-subunit proteins, such as antibodies, the host cell is a yeast, such as Pichia pastoris, and the glycosylated impurity is a glycovariant of the desired recombinant polypeptide, such as an N-linked and/or O-linked glycovariant.
    • 公开了生产和纯化重组蛋白的方法。 特别地,本公开提供了产生和纯化在酵母或丝状真菌细胞中表达的多亚基蛋白质的方法。 监测这些蛋白质的杂质的产生和/或纯化,优选使用凝集素结合分析,以便可以调整一个或多个工艺参数以最大化所需重组蛋白的量并使糖基化杂质的量最小化。 还可以监测过程中是否有其他不需要的产品相关杂质,例如聚集体和核酸。 在示例性实施方案中,重组蛋白质是多亚基蛋白质,例如抗体,宿主细胞是酵母,例如巴斯德毕赤酵母(Pichia pastoris),并且糖基化杂质是所需重组多肽的糖变体,例如N-连接的和/ 或O-连接的糖变体。