Integrated autolysis, DNA hydrolysis and precipitation enables an improved bioprocess for Q-Griffithsin, a broad-spectrum antiviral and clinical-stage anti-COVID-19 candidate.
Integrated autolysis, DNA hydrolysis and precipitation enables an improved bioprocess for Q-Griffithsin, a broad-spectrum antiviral and clinical-stage anti-COVID-19 candidate.
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DOI：
10.1016/j.bej.2022.108403
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发表时间：
2022-04
影响因子：
3.9
通讯作者：
Lynch MD
中科院分区：
文献类型：
--
作者：
Decker JS;Menacho-Melgar R;Lynch MD
整个生物制造行业都需要创新来提高效率和灵活性，特别是在面对 COVID-19 大流行等挑战时。在这里，我们报告了 Q-Griffithsin 的改进生物工艺，Q-Griffithsin 是一种广谱抗病毒药物，目前正在进行 COVID-19 的临床试验。 Q-Griffithsin 在大肠杆菌中以高滴度生产，并纯化至预期的临床级别，无需传统色谱法或任何固定的下游设备。因此，该过程既低成本又高度灵活，有利于降低销售价格和灵活调整产能，这是应对大流行病的两个关键特征。这一过程的简单性得益于一种新颖的单元操作，该操作集成了细胞自溶、核酸自水解和污染物沉淀，基本上完全去除了宿主细胞 DNA，并将宿主细胞蛋白质和内毒素分别减少了 3.6 和 2.4 log10 单位。该单元操作可以在发酵容器中快速进行，从而在发酵后立即获得 100% 产率和 > 99.9% 纯度的 Q-GRFT，并且仅需要流通膜色谱步骤即可进一步去除污染物。使用这种操作或其变体可以改善大肠杆菌中一系列其他高价值蛋白质的生物过程。
Across the biomanufacturing industry, innovations are needed to improve efficiency and flexibility, especially in the face of challenges such as the COVID-19 pandemic. Here we report an improved bioprocess for Q-Griffithsin, a broad-spectrum antiviral currently in clinical trials for COVID-19. Q-Griffithsin is produced at high titer in E. coli and purified to anticipated clinical grade without conventional chromatography or the need for any fixed downstream equipment. The process is thus both low-cost and highly flexible, facilitating low sales prices and agile modifications of production capacity, two key features for pandemic response. The simplicity of this process is enabled by a novel unit operation that integrates cellular autolysis, autohydrolysis of nucleic acids, and contaminant precipitation, giving essentially complete removal of host cell DNA as well as reducing host cell proteins and endotoxin by 3.6 and 2.4 log10 units, respectively. This unit operation can be performed rapidly and in the fermentation vessel, such that Q-GRFT is obtained with 100% yield and > 99.9% purity immediately after fermentation and requires only a flow-through membrane chromatography step for further contaminant removal. Using this operation or variations of it may enable improved bioprocesses for a range of other high-value proteins in E. coli.
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影响因子：
4.6
作者：
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通讯作者：
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影响因子：
2.9
作者：
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通讯作者：
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影响因子：
3.8
作者：
Menacho-Melgar, Romel;Ye, Zhixia;Moreb, Eirik A.;Yang, Tian;Efromson, John P.;Decker, John S.;Wang, Ruixin;Lynch, Michael D.
通讯作者：
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影响因子：
2.2
作者：
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影响因子：
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通讯作者：
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