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首页> 外文期刊>Biotechnology Progress >High-Throughput Ion Exchange Purification of Positively Charged Recombinant Protein in the Presence of Negatively Charged Dextran Sulfate
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High-Throughput Ion Exchange Purification of Positively Charged Recombinant Protein in the Presence of Negatively Charged Dextran Sulfate

机译：存在负电荷硫酸葡聚糖的高通量离子交换纯化正电荷重组蛋白

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摘要

Product quality analyses are critical for developing cell line and bioprocess producing therapeutic proteins with desired critical product quality attributes. To facilitate these analyses, a high-throughput small-scale protein purification (SSP) is required to quickly purify many samples in parallel. Here we develop an SSP using ion exchange resins to purify a positively charged recombinant growth factor PI in the presence of negatively charged dextran sulfate supplemented to improve the cell culture performance. The major challenge in this work is that the strong ionic interaction between PI and dextran sulfate disrupts interaction between PI and chromatography resins. To solve this problem, we develop a two-step SSP using Q Sepharose Fast Flow (QFF) and SP Sepharose XL (SPXL) resins to purify PL The overall yield of this two-step SSP is 78%. Moreover, the SSP does not affect the critical product quality attributes. The SSP was critical for developing the cell line and process producing P1.
机译：产品质量分析对于开发具有所需关键产品质量属性的治疗蛋白的细胞系和生物过程至关重要。为了促进这些分析，需要高通量小规模蛋白质纯化（SSP）来快速并行纯化许多样品。在这里，我们开发了一种使用离子交换树脂的SSP，可在存在负电荷的硫酸右旋糖酐硫酸盐存在的情况下纯化带正电荷的重组生长因子PI，以改善细胞培养性能。这项工作的主要挑战是PI和硫酸葡聚糖之间的强离子相互作用会破坏PI和色谱树脂之间的相互作用。为解决此问题，我们使用Q Sepharose Fast Flow（QFF）和SP Sepharose XL（SPXL）树脂开发了两步SSP，以纯化PL。此两步SSP的总产率为78％。而且，SSP不会影响关键的产品质量属性。 SSP对于开发细胞系和生产P1至关重要。

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