PDF(Pubmed)
Abstract：
At times of pandemics, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the situation demands rapid development and production timelines of safe and effective vaccines for delivering life-saving medications quickly to patients. Typical biologics production relies on using the lengthy and arduous approach of stable single-cell clones. Here, we used an alternative approach, a stable cell pool that takes only weeks to generate compared to a stable single-cell clone that needs several months to complete. We employed the membrane, envelope, and highly immunogenic spike proteins of SARS-CoV-2 to produce virus-like particles (VLPs) using the HEK293-F cell line as a host system with an economical transfection reagent. The cell pool showed the stability of protein expression for more than one month. We demonstrated that the production of SARS-CoV-2 VLPs using this cell pool was scalable up to a stirred-tank 2 L bioreactor in fed-batch mode. The purified VLPs were properly assembled, and their size was consistent with the authentic virus. Our particles were functional as they specifically entered the cell that naturally expresses ACE-2. Notably, this work reports a practical and cost-effective manufacturing platform for scalable SARS-CoV-2 VLPs production and chromatographic purification.
摘要：
在大流行的时候，如严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染，这种情况需要快速开发和生产安全有效的疫苗，以迅速向患者提供挽救生命的药物。典型的生物制品生产依赖于使用稳定的单细胞克隆的漫长而艰巨的方法。这里,我们使用了另一种方法，与需要几个月才能完成的稳定的单细胞克隆相比，仅需数周即可生成的稳定细胞池。我们使用了膜，信封,和SARS-CoV-2的高免疫原性刺突蛋白，使用HEK293-F细胞系作为宿主系统，使用经济的转染试剂生产病毒样颗粒（VLP）。细胞池显示蛋白质表达的稳定性超过一个月。我们证明，使用该细胞池生产SARS-CoV-2VLP可以按补料分批模式扩展到搅拌槽2L生物反应器。纯化的VLP正确组装，它们的大小与真正的病毒一致。我们的颗粒是有功能的，因为它们特别进入自然表达ACE-2的细胞。值得注意的是,这项工作报告了一个实用且具有成本效益的制造平台，用于可扩展的SARS-CoV-2VLP生产和色谱纯化。
